ES2966517T3 - sGC stimulators - Google Patents

Abstract

Compounds of Formulas I' and I are described, which are useful as sGC stimulators, particularly NO-independent heme-dependent stimulators. These compounds are also useful for treating, preventing or managing various disorders described herein. (Automatic translation with Google Translate, without legal value)

Description

DESCRIPTION

sGC stimulators

field of invention

The present description relates to soluble guanylate cyclase (sGC) stimulators, pharmaceutical formulations comprising them and their uses, alone or in combination with one or more additional agents, to treat and/or prevent various diseases, where a increase in the concentration of nitric oxide (NO) or an increase in the concentration of cyclic guanosine monophosphate (cGMP).

Background of the invention

Soluble guanylate cyclase (sGC) is the primary receptor for nitric oxide (NO) in vivo. sGC can be activated by both NO-dependent and NO-independent mechanisms. In response to this activation, the sGC converts GTP to the secondary messenger cyclic GMP (cGMP). The increased level of cGMP, in turn, modulates the activity of downstream effectors, including protein kinases, phosphodiesterases (PDEs), and ion channels.

In the body, NO is synthesized from arginine and oxygen by various nitric oxide synthase (NOS) enzymes and by a sequential reduction of inorganic nitrate. Three distinct isoforms of NOS have been identified: inducible NOS (iNOS or NOSII) in activated macrophage cells; Constitutive neuronal NOS (nNOS or NOSI) involved in neurotransmission and long-term potentiation; and constitutive endothelial NOS (eNOS or NOSIII) which regulates smooth muscle relaxation and blood pressure.

Experimental and clinical evidence indicates that reduced bioavailability and/or responsiveness to endogenously produced NO contributes to the development of cardiovascular, endothelial, renal, and hepatic disease, as well as erectile dysfunction and other sexual disorders (e.g., female sexual or vaginal atrophy). Particularly, the NO signaling pathway is altered in cardiovascular diseases, including, for example, systemic and pulmonary hypertension, heart failure, angina, stroke, thrombosis and other thromboembolic diseases, peripheral arterial diseases, liver fibrosis, lung or kidney and atherosclerosis.

sGC stimulators are also useful in the treatment of lipid-related disorders such as, for example, dyslipidemia, hypercholesterolemia, hypertriglyceridemia, sitosterolemia, fatty liver disease and hepatitis.

Pulmonary hypertension (PH) is a disease characterized by a sustained elevation of blood pressure in the pulmonary vasculature (pulmonary artery, pulmonary vein, and pulmonary capillaries), resulting in right cardiac hypertrophy, ultimately leading to right heart failure and death. In PH, the bioactivity of NO and other vasodilators such as prostacyclin is reduced, while the production of endogenous vasoconstrictors such as endotalin is increased, resulting in excessive pulmonary vasoconstriction. sGC stimulators have been used to treat PH because they promote smooth muscle relaxation, resulting in vasodilation.

Treatment with NO-independent sGC stimulators has also favored the relaxation of smooth muscle in the corpus cavernosum of healthy rabbits, rats and humans, causing penile erection, indicating that sGC stimulators are useful for treating erectile dysfunction.

Heme-dependent, NO-independent sGC stimulators, such as those described herein, have several important differentiating features, including crucial dependence on the presence of the reduced prosthetic heme moiety for their activity, strong synergistic enzyme activation when is combined with NO and stimulation of cGMP synthesis by direct stimulation of sGC, independent of NO. The benzylindazole compound YC-1 was the first sGC stimulator to be identified. Since then, additional sGC stimulators of improved potency and specificity for sGC have been developed. It has been proven that these compounds produce antiaggregant, antiproliferative and vasodilatory effects.

Since compounds that stimulate sGC in a NO-independent manner offer considerable advantages over other current alternative therapies, there is a need to develop novel sGC stimulators. They are potentially useful in the prevention, management and treatment of disorders such as pulmonary hypertension, high blood pressure, heart failure, atherosclerosis, inflammation, thrombosis, fibrosis and renal failure, cirrhosis of the liver, pulmonary fibrosis , erectile dysfunction, female hypoactive sexual desire disorder and vaginal atrophy and other cardiovascular disorders; They are also potentially useful for the prevention, management and treatment of lipid-related disorders.

WO 2013/101830 discloses substituted pyrazoles that are useful as sGC stimulators. WO 2012/003405 discloses compounds that are useful as sGC stimulators.

Summary of the invention

The present invention is directed to compounds represented by the following formulas or pharmaceutically acceptable salts thereof:

The invention is further directed to a pharmaceutical composition comprising the compounds, pharmaceutically acceptable salts thereof, and one or more excipients.

The invention also provides the compounds, or pharmaceutically acceptable salts thereof, for use in the treatment of a disease, condition or health disorder selected from:

(1) peripheral, pulmonary, hepatic, kidney, cardiac or cerebrovascular/endothelial disorders/conditions or diseases otherwise related to circulation selected from:

• disorders related to high blood pressure and reduced coronary blood flow; increased acute and chronic coronary blood pressure; arterial hypertension; vascular disorder due to cardiac and renal complications, heart disease, stroke, cerebral ischemia or renal failure; resistant hypertension; diabetic hypertension; congestive heart failure; diastolic or systolic dysfunction; coronary insufficiency; arrhythmias; reduction of ventricular preload; cardiac hypertrophy; heart/cardiorenal failure syndrome; portal hypertension; endothelial dysfunction or injury;

• thromboembolic disorders and ischemia; myocardial infarction; ictus; transient ischemic attacks (TIA); obstructive thromboanginitis; stable or unstable angina pectoris; coronary spasms; variant angina; Prinzmetal's angina; restenosis resulting from thrombolysis therapies; thrombogenic disorders;

• Alzheimer disease; Parkinson's disease; dementia; vascular cognitive impairment; cerebral vasospasms; head trauma;

• Peripheral arterial disease; occlusive peripheral arterial disease; peripheral vascular disease; hypertonias; Raynaud's syndrome or phenomenon; critical limb ischemia; vasculitis; peripheral embolism; intermittent claudication; vaso-occlusive crises; Duchenne muscular dystrophy; Becker muscular dystrophy; microcirculation abnormalities; problems with exudation or vascular permeability;

• shock; septicemia; cardiogenic shock; control of leukocyte activation; inhibition or modulation of platelet aggregation;

• pulmonary/respiratory conditions; pulmonary hypertension; pulmonary arterial hypertension and associated pulmonary vascular remodeling; localized thrombosis and right cardiac hypertrophy; pulmonary hypertonia; primary pulmonary hypertension; secondary pulmonary hypertension; familial pulmonary hypertension; sporadic pulmonary hypertension, precapillary pulmonary hypertension; idiopathic pulmonary hypertension; thrombotic pulmonary arteriopathy; plexogenic pulmonary arteriopathy; cystic fibrosis; bronchoconstriction or pulmonary bronchoconstriction; acute respiratory distress syndrome; pulmonary fibrosis; lung transplant;

• pulmonary hypertension associated with or related to left ventricular dysfunction, hypoxemia, hypertension of WHO groups I, II, III, IV and V, mitral valve disease, constrictive pericarditis, aortic stenosis, cardiomyopathy, mediastinal fibrosis, pulmonary fibrosis , anomalous pulmonary venous drainage, pulmonary veno-occlusive disease, pulmonary vasculitis, collagen vascular disease, congenital heart disease, pulmonary venous hypertension, interstitial lung disease, sleep disordered breathing, sleep apnea, alveolar hypoventilation disorders, chronic exposure to high altitude, disease pulmonary in neonates, alveolar capillary dysplasia, sickle cell disease; coagulation disorders; chronic thromboembolism, pulmonary embolism due to tumor, parasites or foreign material, connective tissue disease, lupus, schistosomiasis, sarcoidosis, chronic obstructive pulmonary disease, asthma, emphysema, chronic bronchitis, pulmonary capillary hemangiomatosis; histiocytosis

• atherosclerotic diseases or conditions; atherosclerosis; atherosclerosis associated with endothelial injury, platelet and monocyte adhesion and aggregation, smooth muscle proliferation and migration; restenosis; restenosis developed after thrombolysis therapies, percutaneous transluminal angioplasties (PTA), percutaneous transluminal coronary angioplasties (PTCA), and bypasses; inflammation;

• cardiovascular disease associated with metabolic syndrome, obesity, dyslipidemia, diabetes, high blood pressure; lipid-related disorders, such as dyslipidemia, hypercholesterolemia, hypertriglyceridemia, sitosterolemia, fatty liver disease, and hepatitis; preeclampsia; progression of polycystic kidney disease; subcutaneous fat accumulation;

• hepatic cirrhosis; liver cirrhosis associated with chronic liver disease; liver fibrosis, activation of hepatic stellate cells, hepatic fibrous collagen; total collagen accumulation; liver disease of necroinflammatory and/or immunological origin;

• disorders of the urogenital system; kidney fibrosis; kidney failure resulting from chronic kidney disease or failure; renal failure due to tissue accumulation/deposition and injury, progressive sclerosis and glomerulonephritis; prostatic hypertrophy;

• systemic sclerosis;

• cardiac interstitial fibrosis; cardiac remodeling and fibrosis; cardiac hypertrophy;

(2) ischemia or reperfusion injury; ischemia/reperfusion associated with organ transplant, lung transplant, lung transplant, or heart transplant; preservation of blood substituents in trauma patients;

(3) a selected sexual, gynecological, and urological disorder of erectile dysfunction; impotence; premature ejaculation; female sexual dysfunction; female sexual desire dysfunction; hypoactive sexual desire disorder; vaginal atrophy; dyspaneuria; atrophic vaginitis; benign prostatic hyperplasia (BPH) or benign prostatic hypertrophy or enlargement; bladder outlet obstruction; painful bladder syndrome (BPS); interstitial cystitis (IC); overactive bladder syndrome, neurogenic bladder and incontinence; diabetic nephropathy;

(4) ocular diseases or disorders selected from glaucoma, retinopathy, diabetic retinopathy, blepharitis, dry eye syndrome, Sjogren's syndrome;

(5) hearing diseases or disorders selected from hearing impairment; partial or total hearing loss; partial or total deafness; tinnitus; noise-induced hearing loss;

(6) selected topical or skin disorders or conditions of dermal fibrosis, scleroderma, skin fibrosis;

(7) wound healing; wound healing in diabetics; improvement of microvascular perfusion; improving microvascular perfusion after injury, to counteract the inflammatory response in perioperative care; anal fissures; diabetic ulcers; and

(8) other diseases or conditions selected from cancer metastasis; osteoporosis, gastroparesis; functional dyspepsia; diabetic complications; diseases associated with endothelial dysfunction; and neurological disorders associated with decreased nitric oxide production.

Detailed description of the invention

Reference will now be made in detail to certain embodiments of the invention, examples of which are illustrated in the attached structures and formulas. While the invention will be described in conjunction with the listed embodiments, it will be understood that these are not intended to limit the invention to those embodiments. Rather, the invention is intended to cover all alternatives, modifications and equivalents that may be included within the scope of the present invention as defined in the embodiments. The present invention is not limited to the methods and materials described herein, but includes any methods and materials similar or equivalent to those described herein that could be used in the practice of the present invention. In the event that one or more of the literature references, patents or similar materials differ from or contradict this application, including, but not limited to, defined terms, use of terms, described techniques or the like, the indicated shall prevail. in this application.

Definitions and general terminology

For the purposes of this description, chemical elements are identified according to the periodic table of the elements, CAS version and the Handbook of Chemistry and Physics, 75th ed. 1994. Additionally, general principles of organic chemistry are described in “Organic Chemistry”, Thomas Sorrell, University Science Books, Sausalito: 1999, and “March's Advanced Organic Chemistry”, 5th ed., Smith, M. B. and March, J. , eds. John Wiley & Sons, New York: 2001.

As used herein, the expression “to” refers to zero or any integer equal to or less than the number following the expression. For example, "up to 3" means any one of 0, 1, 2, or 3. As described herein, a specified numerical range of atoms includes any integer within the range. For example, a group that has 1 to 4 atoms could have 1, 2, 3, or 4 atoms. When any variable appears more than once in any position, its definition in each case is independent of the other cases.

The selection of substituents and combinations contemplated by this description are only those that give rise to the formation of stable or chemically viable compounds. These choices and combinations will be apparent to one skilled in the art and can be determined without undue experimentation. As used herein, the term "stable" refers to compounds that are not substantially altered when subjected to conditions that permit their production, detection and, in some embodiments, their recovery, purification, and use for one or more purposes. for the purposes described in this description. In some embodiments, a stable compound is one that is not substantially altered when maintained at a temperature of 25°C or less, in the absence of humidity or other chemically reactive conditions, for at least one week. A chemically viable compound is a compound that can be prepared by a person skilled in the art based on the descriptions offered herein supplemented, if necessary, with relevant knowledge of the art.

A compound, such as the compounds of claims 1 and 2, may be present in its free form (for example, an amorphous form, or a crystalline form or a polymorph). Under certain conditions, compounds can also form coforms. As used herein, the term coform is synonymous with the term multicomponent crystalline form. When one of the components in the coform has clearly transferred a proton to the other component, the resulting coform is called a “salt.” The formation of a salt is determined by how large the difference in pKas is between the elements that make up the mixture. For the purposes of this description, compounds include pharmaceutically acceptable salts, although the term “pharmaceutically acceptable salts” is not explicitly stated.

Unless only one of the isomers is specifically drawn or named, the structures depicted herein also include all stereoisomeric forms (e.g., enantiomeric, diastereomeric, atropoisomeric, and cis-trans isomeric forms) of the structure; for example, the Ry S configurations for each asymmetric center, the RaySa configurations for each asymmetric axis, the double link configurations

(Z) and (E) and the cis and trans conformational isomers. Therefore, simple stereochemical isomers, as well as racemates, and mixtures of enantiomers, diastereomers and cis-trans (double bond or conformational) isomers of the present compounds are within the scope of this description. Unless otherwise indicated, all tautomeric forms of the compounds of the present description are also within the scope of the invention. As an example, a substituent drawn as follows:

where R can be hydrogen, would include both compounds shown below:

The present description also covers isotopically labeled compounds that are identical to those mentioned in the present description, except that one or more atoms are replaced by an atom having an atomic mass or mass number different from the atomic mass or mass number. usually found in nature. All isotopes of any particular atom or element as specified are contemplated within the scope of the compounds of the invention and their uses. Illustrative isotopes that can be incorporated into the compounds of the invention include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, sulfur, fluorine, chlorine, and iodine, such as 2H, 3H, 11C, 13C, 14C, 13N, 15N, 15O, 17O, 18O, 32P, 33P, 35S, 18F, Certain isotopically labeled compounds of the present invention (for example, those labeled with 3H and

14C) are useful in distribution assays of compounds and/or substrate tissues. Tritiated isotopes (i.e.

3H) and carbon-14 (i.e. 14C) are useful because of their easy preparation and detectability. Furthermore, replacement with heavier isotopes such as deuterium (i.e., 2H) may offer certain therapeutic advantages due to greater metabolic stability (e.g., longer in vivo half-life or lower dosage requirements) and therefore may preferred in some circumstances. Positron-emitting isotopes, such as 15O, 13N, 11C,

and<18>F are useful for positron emission tomography (PET) studies to examine substrate receptor occupancy. The isotopically labeled compounds of the present invention can be prepared, generally, by the following procedures analogous to those described in the schemes and/or in the examples included below, substituting an isotopically labeled reagent for a non-isotopically labeled reagent.

As used herein, the term “aliphatic” or “aliphatic group” means a straight-chain (i.e., unbranched) or branched, substituted or unsubstituted hydrocarbon chain that is fully saturated or that contains one or more units. of unsaturation. Unless otherwise indicated, aliphatic groups contain 1 20 aliphatic carbon atoms. In some embodiments, the aliphatic groups contain from 1 to 10 aliphatic carbon atoms. In other embodiments, the aliphatic groups contain from 1 to 8 aliphatic carbon atoms. In other embodiments, the aliphatic groups contain from 1 to 6 aliphatic carbon atoms. In other embodiments, the aliphatic groups contain 1-4 aliphatic carbon atoms, and in other embodiments, the aliphatic groups contain 1-3 aliphatic carbon atoms. Suitable aliphatic groups include, but are not limited to, substituted or unsubstituted linear or branched alkyl, alkenyl or alkynyl groups. Specific examples of aliphatic groups include, but are not limited to: methyl, ethyl, propyl, butyl, isopropyl, isobutyl, vinyl, sec-butyl, ferf-butyl, butenyl, propargyl, acetylene and the like. For complete clarity, the term “aliphatic chain” can be used interchangeably with the term “aliphatic” or “aliphatic group”.

As used herein, the term “alkyl” refers to a saturated straight-chain or branched monovalent hydrocarbon radical. Unless otherwise noted, an alkyl group contains 1-20 carbon atoms (e.g., 1-20 carbon atoms, 1-10 carbon atoms, 1-8 carbon atoms, 1-6 carbon atoms, 1 -4 carbon atoms or 1-3 carbon atoms). Examples of alkyl groups include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, s-butyl, t-butyl, pentyl, hexyl, heptyl, octyl and the like.

The term “alkenyl” refers to a straight-chain or branched monovalent hydrocarbon radical with at least one site of unsaturation, i.e., an sp<2>carbon-carbon double bond, wherein the alkenyl radical includes radicals having “alkenyl” orientations. cis” and “trans” or, alternatively, “E” and “Z” orientations. Unless otherwise noted, an alkenyl group contains 2-20 carbon atoms (e.g., 2-20 carbon atoms, 2-10 carbon atoms, 2-8 carbon atoms, 2-6 carbon atoms, 2 -4 carbon atoms or 2-3 carbon atoms). Examples include, but are not limited to, vinyl, allyl and the like.

The term "alkynyl" refers to a linear or branched monovalent hydrocarbon radical with at least one unsaturation site, that is, a sp carbon-carbon triple bond. Unless otherwise noted, an alkynyl group contains 2-20 carbon atoms (e.g., 2-20 carbon atoms, 2-10 carbon atoms, 2-8 carbon atoms, 2-6 carbon atoms, 2 -4 carbon atoms or 2-3 carbon atoms). Examples include, but are not limited to, ethinyl, propynyl and the like.

The term “carbocyclic” refers to a ring system made up of only carbon and hydrogen atoms. Unless otherwise indicated, throughout this description, carbocycle is used synonymously with “non-aromatic carbocycle” or “cycloaliphatic”. In some cases, the term may be used in the expression “aromatic carbocycle” and in this case refers to an “aryl group”, as defined below.

The term "cycloaliphatic" (or "non-aromatic carbocycle", "non-aromatic carbocyclyl", "non-aromatic carbocyclic") refers to a cyclic hydrocarbon that is completely saturated or that contains one or more unsaturation units but that is not aromatic, and that it has a single point of attachment to the rest of the molecule. Unless otherwise noted, a cycloaliphatic group may be monocyclic, bicyclic, tricyclic, fused, spiro, or bridged. In one embodiment, the term "cycloaliphatic" refers to a C<3>-C<12>monocyclic hydrocarbon or a C<7>-C<12>bicyclic hydrocarbon. In some embodiments, any individual ring in a bicyclic or tricyclic ring system has 3-7 members. Suitable cycloaliphatic groups include, but are not limited to, cycloalkyl, cycloalkenyl and cycloalkynyl. Examples of aliphatic groups include cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cycloheptyl, cycloheptenyl, norbornyl, cyclooctyl, cyclononyl, cyclodecyl, cycloundecyl, cyclododecyl and the like.

The term “cycloaliphatic” also includes polycyclic ring systems in which the non-aromatic carbocyclic ring may be “fused” to one or more aromatic or non-aromatic carbocyclic or heterocyclic rings or combinations thereof, provided that the radical or point of attachment is on the non-aromatic carbocyclic ring.

As used herein, “cycloalkyl” refers to a ring system in which it is completely saturated and which has a single point of attachment to the rest of the molecule. Unless otherwise noted, a cycloalkyl group may be monocyclic, bicyclic, tricyclic, fused, spiro, or bridged. In one embodiment, the term "cycloalkyl" refers to a saturated C<3>-C<12>monocyclic hydrocarbon or a saturated C<7>-C<12>bicyclic hydrocarbon. In some embodiments, any individual ring in a bicyclic or tricyclic ring system has 3-7 members. Suitable cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cycloheptenyl, norbornyl, cyclooctyl, cyclononyl, cyclodecyl, cycloundecyl, cyclododecyl and the like.

As used herein, "heterocycle" (or "heterocycle" or "heterocyclic") refers to a ring system in which one or more ring members is an independently selected heteroatom, which is completely saturated or which contains a or more unsaturation units, but is not aromatic, and has a single point of attachment to the rest of the molecule. Unless otherwise indicated, throughout this description, heterocycle is used synonymously with “non-aromatic heterocycle”. In some cases, the term may be used in the expression "aromatic heterocycle" and in this case refers to a "heteroaryl group", as defined below. The term heterocycle also includes fused, spiro or bridged heterocyclic ring systems. Unless otherwise indicated, a heterocycle may be monocyclic, bicyclic or tricyclic. In some embodiments, the heterocycle has 3-18 ring members wherein one or more ring members is a heteroatom independently selected from oxygen, sulfur or nitrogen, and each ring of the system contains 3 to 7 ring members. In other embodiments, a heterocycle may be a monocycle having 3-7 ring members (2-6 carbon atoms and 1-4 heteroatoms) or a bicycle having 7-10 ring members (4-9 carbon atoms). and 1-6 heteroatoms). Examples of bicyclic heterocyclic ring systems include, but are not limited to: adamantanyl, 2-oxa-bicyclo[2.2.2]octyl, 1-aza-bicyclo[2.2.2]octyl.

As used herein, the term “heterocycle” also includes polycyclic ring systems, wherein the heterocyclic ring is fused to one or more aromatic or non-aromatic carbocyclic or heterocyclic rings, or combinations thereof, provided that the radical or attachment point is on the heterocyclic ring.

Examples of heterocyclic rings include, but are not limited to, the following monocycles: 2-tetrahydrofuranyl, 3-tetrahydrofuranyl, 2-tetrahydrothiophenyl, 3-tetrahydrothiophenyl, 2-morpholino, 3-morpholino, 4-morpholino, 2-thiomorpholino, 3- Tiomorfolino, 4-iTiomorfolino, 1 -Pyrolidinilo, 2-Pyrolidinilo, 3-Pyrolidinilo, 1 -Tetrahydropiperazinil olinyl, 4-pyrazolidinyl, 5-pyrazolidinyl, 1-piperidinyl, 2-piperidinyl, 3-piperidinyl, 4-piperidinyl, 2-thiazolidinyl, 3-thiazolidinyl, 4-thiazolidinyl, 1-imidazolidinyl, 2-imidazolidinyl, 4-imidazolidinyl, 5- imidazolidinyl; and the following bicycles: 3-1H-benzimidazol-2-one, 3-(1-alkyl)-benzimidazol-2-one, indolinyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl, benzothiolane, benzodithiane, and 1,3-dihydro-imidazole-2- ona.

As used herein, the term “aryl” (as in “aryl ring” or “aryl group”), used alone or as part of a larger moiety, as in “aralkyl,” “aralkoxy,” “aryloxyalkyl.” ", refers to a carbocyclic ring system where at least one ring in the system is aromatic and has a single point of attachment to the rest of the molecule. Unless otherwise indicated, an aryl group may be monocyclic, bicyclic or tricyclic and contain 6-18 ring members. The term also includes polycyclic ring systems, where the aryl ring is fused to one or more aromatic or non-aromatic carbocyclic or heterocyclic rings, or combinations thereof, provided that the radical or point of attachment is on the aryl ring. Examples of aryl rings include, but are not limited to, phenyl, naphthyl, indanyl, indenyl, tetralin, fluorenyl, and anthracenyl.

The term "aralkyl" refers to a radical that has an aryl ring substituted with an alkylene group, where the open end of the alkylene group allows the aralkyl radical to bond to another part of the compound. The alkylene group is a bivalent, straight or branched chain, saturated hydrocarbon group. As used herein, the term “C<7-12>aralkyl” means an aralkyl radical wherein the total number of carbon atoms in the aryl ring and the combined alkylene group is 7 to 12. Examples of “aralkyl” include, but are not limited to, a phenyl ring substituted by a C<1-6>alkylene group, for example, benzyl and phenylethyl, and a naphthyl group substituted by a C<1-2>alkylene group.

The term “heteroaryl” (or “heteroaromatic” or “heteroaryl group” or “aromatic heterocycle”) used alone or as part of a larger moiety, as in “heteroaralkyl” or “heteroarylalkoxy” refers to a ring system where At least one ring in the system is aromatic and contains one or more heteroatoms, where each ring in the system contains 3 to 7 ring members and has a single point of attachment to the rest of the molecule. Unless otherwise noted, a heteroaryl ring system may be monocyclic, bicyclic, or tricyclic and have a total of five to fourteen ring members. In one embodiment all rings in a heteroaryl system are aromatic. Also included in this definition are heteroaryl radicals where the heteroaryl ring is fused with one or more aromatic or non-aromatic carbocyclic or heterocyclic rings, or combinations thereof, provided that the radical or point of attachment is on the heteroaryl ring. As used herein, the bicyclic heteroaromatic system 6,5, for example, is a six-membered heteroaromatic ring fused to a second five-membered ring, wherein the radical or attachment point is on the six-membered ring. .

Heteroaryl rings include, but are not limited to, the following monocycles: 2-furanyl, 3-furanyl, N-imidazolyl, 2-imidazolyl, 4-imidazolyl, 5-imidazolyl, 3-isoxazolyl, 4-isoxazolyl, 5-isoxazolyl , 2-oxazolyl, 4-oxazolyl, 5-oxazolyl, N-pyrrolyl, 2-pyrrolyl, 3-pyrrolyl, 2-pyridyl, 3-pyridyl, 4-pyridyl, 2-pyrimidinyl, 4-pyrimidinyl, 5-pyrimidinyl, pyridazinyl (e.g. 3-pyridazinyl), 2-thiazolyl, 4-thiazolyl, 5-thiazolyl, tetrazolyl (e.g. 5-tetrazolyl), triazolyl (e.g. 2-triazolyl and 5-triazolyl), 2-thienyl, 3 -thienyl, pyrazolyl (e.g. 2-pyrazolyl), isothiazolyl, 1,2,3-oxadiazolyl, 1,2,5-oxadiazolyl, 1,2,4-oxadiazolyl, 1,2,3-triazolyl, 1,2 ,3-thiadiazolyl, 1,3,4-thiadiazolyl, 1,2,5-thiadiazolyl, pyrazinyl, 1,3,5-triazinyl, and the following bicycles: benzimidazolyl, benzofuryl, benzothiophenyl, benzopyrazinyl, benzopyranonyl, indolyl (e.g. , 2-indolyl), purinyl, quinolinyl (for example, 2-quinolinyl, 3-quinolinyl, 4-quinolinyl) and isoquinolinyl (for example, 1-isoquinolinyl, 3-isoquinolinyl or 4-isoquinolinyl).

As used herein, "cycle" (or "cyclic", or "cyclic moiety") encompasses monocyclic, bicyclic and tricyclic ring systems including cycloaliphatic, heterocyclic, aryl or heteroaryl, each of which has been defined previously.

“Fused” bicyclic ring systems comprise two rings that share two adjacent ring atoms.

“Bridged” bicyclic ring systems comprise two rings that share three or four adjacent ring atoms. As used herein, the term “bridge” refers to an atom or a chain of atoms that connect two different parts of a molecule. The two atoms that are connected through the bridge (usually, but not always, two tertiary carbon atoms) are called “bridgeheads.” In addition to the bridge, the two bridgeheads are connected by at least two individual atoms or chains of atoms. Examples of bridging bicyclic ring systems include, but are not limited to, adamantanyl, norbornanyl, bicyclo[3.2.1]octyl, bicyclo[2.2.2]nonyl, bicyclo[3.3.1]nonyl, bicyclo[3.2.3] ]nonyl, 2-oxa-bicyclo[2.2.2]octyl, 1-aza-bicyclo[2.2.2]octyl, 3-aza-bicyclo[3.2.1]octyl, and 2,6-dioxa-tricyclo[3.3. 1.03,7]nonyl. “Spiro” bicyclic ring systems share only one ring atom (usually a quaternary carbon atom) between the two rings.

The term "ring atom" refers to an atom such as C, N, O or S that forms part of the ring of an aromatic ring, a cycloaliphatic ring, a heterocyclic ring or a heteroaryl ring. A “substitutable ring atom” is a ring carbon or nitrogen atom bonded to at least one hydrogen atom. The hydrogen can optionally be replaced with a suitable substituent group. Therefore, the term “substitutable ring atom” does not include ring nitrogen or carbon atoms that are shared when two rings are fused. Furthermore, “substitutable ring atom” does not include carbon atoms or nitrogen atoms when the structure describes that they are already attached to one or more radicals other than hydrogen and no hydrogens are available for substitution.

"Heteroatom" means one or more of oxygen, sulfur, nitrogen, phosphorus, or silicon, including any oxidized form of nitrogen, sulfur, phosphorus, or silicon, the quaternized form of any basic nitrogen, or a substitutable nitrogen of a heterocyclic ring or heteroaryl, for example N (as in 3,4-dihydro-2H-pyrrolyl), NH (as in pyrrolidinyl) or NR+ (as in N-substituted pyrrolidinyl).

Two independent cases of a variable can be taken together with the atom(s) to which each variable is attached to form a 5-8 membered heterocyclic, aryl or cycloaliphatic heteroaryl ring or a 3-membered cycloaliphatic ring. -8 members. Illustrative rings that are formed when two independent elements of a substituent are taken together with the atom(s) to which each variable is attached include, but are not limited to, the following: a) two independent elements of a substituent that are attached to the same atom and taken together with that atom to form a ring, where both elements of the substituent are taken together with the atom to which they are attached to form a heterocyclyl, heteroaryl, cycloaliphatic or aryl ring, where the group is attached to the rest of the molecule through a single binding point; and b) two independent elements of a substituent that are attached to different atoms and are taken together with both atoms to form a heterocyclyl, heteroaryl, cycloaliphatic or aryl ring, where the ring that is formed has two points of attachment with the rest of the molecule. For example, when a phenyl group is substituted with two -ORo elements in formula D1:

These two -ORo elements are taken together with the carbon atoms to which they are attached to form a fused 6-membered oxygen-containing ring as in formula 2:

It will be appreciated that a number of different rings can be formed when two independent elements of a substituent are taken together with the atom(s) to which each substituent is attached and that the examples detailed above should not be considered limiting.

An alkyl or aliphatic chain may optionally be interrupted with another atom or group. This means that a methylene unit of the alkyl or aliphatic chain may optionally be substituted with said other atom or group. Unless otherwise noted, optional replacements form a chemically stable compound. Optional interrupts can occur both within the chain and at the end of the chain; that is, at the point of attachment or attachments to the rest of the molecule and/or at the terminal end. Furthermore, two optional replacements can be adjacent to each other within a chain as long as a chemically stable compound is obtained.

Unless otherwise indicated, if the replacement or interruption occurs at a terminal end of the chain, the replacement atom is bonded to an H atom at the terminal end. For example, if -CH<2>CH<2>CH<3> were optionally interrupted with -O-, the resulting compound could be -OCH<2>CH<3>, -CH<2>OCH<3>or -CH<2>CH<2>OH. As another example, if the divalent linking group -CH<2>CH<2>CH<2> were optionally interrupted with -O-, the resulting compound could be -OCH<2>CH<2>-, -CH< 2>OCH<2>- or -CH<2>CH<2>O-. Optional replacements can also completely replace all carbon atoms in a chain. For example, an aliphatic C<3>could be completely replaced by -N(R')-, -C(O)- and -N(R')- to form -N(R')C(O)N( R')- (a urea).

In general, the term "neighboring" refers to the placement of substituents in a group that includes two or more carbon atoms, where the substituents are bonded to adjacent carbon atoms.

In general, the term "geminal" refers to the placement of substituents in a group that includes two or more carbon atoms, where the substituents are attached to the same carbon atom.

The terms “terminally” and “internally” refer to the location of a group within a hydrocarbyl substituent. A group is terminal when the group is present at the end of the substituent that is not additionally attached to the rest of the chemical structure. Carboxyalkyl, i.e. R<X>O(O)C-alkyl is an example of a terminally used carboxyl group. A group is internal when the group is present at the center of a substituent at the end of the substituent attached to the rest of the chemical structure. Alkylcarboxyl (e.g., alkyl-C(O)O- or alkyl-O(CO)-) and alkylcarboxyaryl (e.g., alkyl-C(O)O-aryl or alkyl-O(CO)-aryl-) are examples. of carboxyl groups used internally.

As described herein, a traced bond of a substituent to the center of a ring within a multiple ring system (as shown below), represents the substitution of the substituent at any substitutable position on any of the rings within the multiple ring system. For example, formula D3 represents a possible substitution in any of the positions shown in formula D4:

This also applies to multiple ring systems fused to optional ring systems (which would be represented by dashed lines). For example, in formula D5, X is an optional substituent for both an A ring and a B ring.

If, however, two rings in a multiple ring system each have different substituents drawn from the center of each ring, then, unless otherwise noted, each substituent only represents substitution on the ring to which it is attached. For example, in formula D6, Y is an optional substituent for ring A only, and X is an optional substituent for ring B only.

As used herein, the terms “alkoxy” and “alkylthio” refer to an alkyl group, as defined above, attached to the molecule, or to another chain or ring, through an oxygen atom (“ alkoxy” i.e. -O-alkyl) or sulfur (“alkylthio”, i.e. -S-alkyl).

The terms Cn-m “alkoxyalkyl”, Cn-m “alkoxyalkenyl”, Cn-m “alkoxyaliphatic”, and Cn-m “alkoxyalkoxy” mean alkyl, alkenyl, aliphatic or alkoxy, as the case may be, substituted with one or more alkoxy groups, wherein the total combined number of carbons of the alkyl and alkoxy, alkenyl and alkoxy groups, aliphatic groups and alkoxy groups or alkoxy and alkoxy groups, combined, as the case may be, is between the values of n and m. For example, a C<4-6>alkoxyalkyl has a total of 4-6 carbons divided between the alkyl and alkoxy parts; p. e.g. -can be -CH<2>OCH<2>CH<2>CH<3>, -CH<2>CH<2>OCH<2>CH<3>or -CH<2>CH<2>CH< 2>OCH<3>.

When the moieties described in the previous paragraph are optionally substituted, they may be substituted in one or both of the parts on either side of the oxygen or sulfur. For example, an optionally substituted C<4>alkoxyalkyl could be, for example, -CH<2>CH<2>OCH<2>(Me)CH<3>or -CH<2>(OH)O CH<2 >CH<2>CH<3>; a C<5>alkoxyalkenyl could be, for example, -CH=CHO CH<2>CH<2>CH<3>or -CH=CHCH<2>OCH<2>CH<3>.

The terms aryloxy, arylthio, benzyloxy or benzylthio refer to an aryl or benzyl group attached to the molecule, or to another chain or ring, through an oxygen atom ("aryloxy", benzyloxy, for example, -O-Ph , -OCH<2>Ph) or sulfur (“arylthio”, for example, -S-Ph, -S-CH<2>Ph). Furthermore, the terms "aryloxyalkyl", "benzyloxyalkyl", "aryloxyalkenyl" and "aryloxyaliphatic" mean alkyl, alkenyl or aliphatic, as the case may be, substituted with one or more aryloxy or benzyloxy groups, as the case may be. In this case, the number of atoms for each aryl, aryloxy, alkyl, alkenyl or aliphatic will be indicated separately. Therefore, a 5-6-membered aryloxy(C<1-4>alkyl) is a 5-6-membered aryl ring, linked via an oxygen atom to a C<1-4>alkyl chain that, in turn, Instead, it is linked to the rest of the molecule through the terminal carbon of the C<1-4> alkyl chain.

As used herein, the terms “halogen” or “halo” mean F, Cl, Br or I.

The terms "haloalkyl", "haloalkenyl", "haloaliphatic" and "haloalkoxy" mean alkyl, alkenyl, aliphatic or alkoxy, as the case may be, substituted with one or more halogen atoms. For example, a C<1-3>haloalkyl could be -CFHCH<2>CHF<2>and a C<1-2>haloalkoxy could be -OC(Br)HCHF<2>. This term includes perfluorinated alkyl groups, such as -C F<3>and -CF<2>CF<3>.

As used herein, the term “cyano” refers to -CN or -CeN.

The terms "cyanoalkyl", "cyanoalkenyl", "cyanoaliphatic", and "cyanoalkoxy" mean alkyl, alkenyl, aliphatic or alkoxy, as the case may be, substituted with one or more cyano groups. For example, a C<1-3>cyanoalkyl could be -C(CN)<2>CH<2>CH<3>and a C<1-2>cyanoalkenyl could be =CHC(CN)H<2>.

As used herein, an “amino” group refers to -NH<2>.

The terms "aminoalkyl", "aminoalkenyl", "aminoaliphatic" and "aminoalkoxy" mean alkyl, alkenyl, aliphatic or alkoxy, as the case may be, substituted with one or more amino groups. For example a C<1-3>aminoalkyl could be -CH(NH<2>)CH<2>CH<2>NH<2>and a C<1-2>aminoalkoxy could be -OCH<2>CH< 2>NH<2>.

The term “hydroxyl” or “hydroxy” refers to -OH.

The terms "hydroxyalkyl", "hydroxyalkenyl", "hydroxyaliphatic" and "hydroxyalkoxy" mean alkyl, alkenyl, aliphatic or alkoxy, as the case may be, substituted with one or more -OH groups. For example, a C<1-3>hydroxyalkyl could be -CH<2>(CH<2>OH)CH<3>and a C<4>hydroxyalkoxy could be -OCH<2>C(CH<3>) (OH)CH<3>.

As used herein, a "carbonyl", used alone or in connection with another group refers to -C(O) -or -C(O)H. For example, as used herein, an "alkoxycarbonyl" refers to a group such as -C(O)O(alkyl).

As used herein, an "oxo" refers to =O, where the oxo is usually, but not always, bonded to a carbon atom (for example, it may also be bonded to a sulfur atom). An aliphatic chain may be optionally interrupted by a carbonyl group or may be optionally substituted by an oxo group, and both expressions refer to the same thing: for example, -CH<2>-C(O)-CH<3>.

As used herein in the context of resin chemistry (e.g., using solid resins or soluble resins or beads), the term “linking group” refers to a bifunctional chemical moiety that links a compound to a solid support or soluble support.

As used herein, in all other situations, a “linking group” refers to a divalent group in which the two free valences are on different atoms (e.g., carbon or heteroatom) or are in the same atom but can be substituted by two different substituents. For example, a methylene group may be a C<1>(-CH<2>-) alkyl linking group that may be substituted by two different groups, one for each of the free valences (e.g., as in Ph- CH<2>-Ph, where methylene acts as a linking group between two phenyl rings). Ethylene may be a C<2>(-CH<2>CH<2>-) alkyl linking group where the two free valences are on different atoms. The amide group, for example, can act as a linking group when placed at an internal position in a chain (e.g., -CONH-). A linking group can be the result of interrupting an aliphatic chain by certain functional groups or replacing the methylene units in said chain with said functional groups. E.g., a linking group may be a C<1-6>aliphatic chain in which up to two methylene units are substituted by -C(O)- or -N H - (as in -C H<2>- NH-CH<2>-C(O)-CH<2>- or - CH<2>-NH-C(O)-CH<2>-). An alternative way to define the same group -CH<2>-NH-CH<2>-C(O)-CH<2>- and - CH<2>-NH-C(O)-CH<2>- is a C3 alkyl chain optionally interrupted by up to two -C(O) - or -N H - residues. Cyclic groups can also form linking groups: for example, a 1,6-cyclohexanediyl can be a linking group between two R groups, as in

A linking group may be additionally substituted at any part or position.

Divalent groups of the type R-CH= or R2C= are also possible, where the two free valences are on the same atom and are attached to the same substituent. In this case, they will be referred to by their accepted IUPAC names. <For example, an alkylidene (such as, for example, a methylidene (=CH2) or an ethylidene>(=C<h CH3)) would not be>included by the definition of a link group in this description.

As used herein, the term “protecting group” refers to an agent used to temporarily block one or more desired reactive sites on a multifunctional compound. In certain embodiments, a protecting group has one or more, or preferably all, of the following characteristics: a) it reacts, selectively, in good yield giving rise to a protected substrate that is stable to reactions that occur in one or more of the other reactive sites; and b) it is selectively extractable with good yield using reagents that do not attack the regenerated functional group. Illustrative protecting groups are detailed in Greene, T. W. et al., “Protective Groups in Organic Synthesis”, Third Edition, John Wiley & Sons, New York: 1999. As used herein, the term “nitrogen protecting group” ” refers to an agent used to temporarily block one or more desired reactive nitrogen sites in a multifunctional compound. Preferred nitrogen protecting groups also possess the characteristics illustrated above, and certain illustrative nitrogen protecting groups are detailed in Chapter 7 in Greene, T. W., Wuts, P. G, “Protective Groups in Organic Synthesis”, Third Edition, John Wiley & Sons, New York: 1999.

As used herein, the term “displaceable moiety” or “leaving group” refers to a group that is associated with an aliphatic or aromatic group, as used herein, and can be displaced by nucleophilic attack. by a nucleophile.

As used herein, “amide coupling agent” or “amide coupling reagent” means a compound that reacts with the hydroxyl moiety of a carboxyl moiety, thereby rendering it susceptible to nucleophilic attack. Illustrative amide coupling agents include DIC (diisopropylcarbodiimide), EDCI (1-ethyl-3-(3-dimethylaminopropyl)carbodiimide), DCC (dicyclohexylcarbodiimide), BOP (benzotriazol-1-yloxy-tris(dimethylamino)-phosphonium hexafluorophosphate), pyBOP ((benzotriazol-1-yloxy)tripyrrolidinophosphonium hexafluorophosphate), etc.

The compounds of the invention are defined herein by their chemical structures and/or chemical names. When a compound is designated by both a chemical structure and a chemical name, and the chemical structure and chemical name conflict, the chemical structure is decisive as to the identity of the compound.

Composite embodiments

The invention relates to a compound represented by the following formulas, or pharmaceutically acceptable salts thereof:

Tables 1A, Table 1B, Table 1C and Table 1D list the claimed compounds of the invention (compounds 1-405 and 1-530), and also disclose compounds I-1 to I-404, I-406 to 1-529 , and 1-531 to I-634, which are not claimed.

Table 1A

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Compound preparation methods

The compound of the present invention can be prepared according to the schemes and examples represented and described below. Unless otherwise indicated, the starting materials and the different intermediate products can be obtained from commercial sources, prepared from commercial compounds or prepared using well-known synthetic methods. A process for preparing the compounds as disclosed herein is described, but not claimed.

General synthetic procedures for the compound of this invention are described below. The synthetic schemes are presented as examples and do not limit the scope of the invention in any way.

General procedure A

Dione enolate formation: To a solution of ketone A in THF cooled to -78 °C, LiHMDS (e.g., 0.9 equiv, 1.0 M in toluene) was added dropwise via syringe. The reaction was allowed to warm to 0 °C, then charged with diethyl oxalate (1.2 equiv). At that time, the reaction was warmed to room temperature and stirred at this temperature until it was considered complete (e.g., using TLC or LC/MS analysis). After completion of the reaction (reaction time was typically 45 minutes), the dione enolate B product was used “as is” in step 2, i.e., the cyclization step, without further purification.

Stage 2:

Pyrazole formation: Dione enolate B was diluted with ethanol and consecutively loaded with HCl (e.g., 3 equiv., 1.25 M solution in ethanol) and arylhydrazine hydrate (e.g., 1.15 equiv.). The reaction mixture was heated to 70°C and stirred at this temperature until cyclization was considered complete (e.g., by LC/Ms analysis, typically 30 minutes). Once complete, the reaction mixture was carefully treated with solid sodium bicarbonate (e.g., 4 equiv.) and diluted with dichloromethane and water. The layers were separated and the aqueous layer was further diluted with water before extraction with dichloromethane (3 times). The combined organic layers were washed with brine, dried over MgSO4, filtered, and concentrated in vacuo. The resulting pyrazole C was then purified by SiO2 chromatography using an appropriate gradient of EtOAc in hexanes.

Stage 3:

Formation of amidine: To a suspension of NH4Cl (e.g., 5 equiv.) in toluene cooled to 0 °C, AlMe3 (e.g., 5 equiv., 2.0 M solution in toluene) was added dropwise with a syringe. . The reaction was allowed to warm to room temperature and stirred at this temperature until no further bubbling was observed. Pyrazole C was added in 1 part to the reaction mixture, heated to 110 °C, and stirred at this temperature until considered complete (e.g., using TLC or LC/MS analysis). Once complete, the reaction was cooled, treated with excess methanol, and stirred vigorously for 1 hour at room temperature. The slurry was filtered, and the resulting solid cake was washed with methanol. The filtrate was concentrated in vacuo and the resulting solids were resuspended in a solvent mixture of ethyl acetate:isopropyl alcohol = 5:1. The reaction was further treated with saturated sodium carbonate solution and stirred for 10 minutes before separating the layers. The aqueous layer was extracted with the solvent mixture of ethyl acetate:isopropyl alcohol = 5:1 (3 times), and the combined organic materials were washed with brine. The organic substances were further dried over MgSO4, filtered, and the solvent was removed in vacuo. The resulting amidine D was used as is in subsequent steps without further purification.

Stage 4:

Formation of pyrimidone:Amidine D was suspended in ethanol and shaken vigorously at 23 °C to promote complete solvation. The reaction was further treated with sodium 3-ethoxy-2-fluoro-3-oxoprop-1-en-1-olate (e.g., 3 equiv.), and the flask was equipped with a reflux condenser. The reaction was placed in a preheated oil bath maintained at 90 °C and stirred until full consumption of starting material was observed by LC/MS (reaction times were typically 1 h). The contents were cooled to 23 °C, and the reaction mixture was acidified with HCl (e.g., 3 equiv., 1.25 ml solution in EtOH). The mixture was stirred for 30 minutes and most of the solvent was removed in vacuo. The contents were resuspended in ether and water (1:1 mixture), and the resulting suspension was stirred for 20 min. The suspension was filtered under vacuum, and the solid cake was rinsed with additional water and ether and dried under high vacuum overnight. The resulting pyrimidone E was used as is in subsequent steps without further purification.

General procedure B

A solution of amino-type nucleophile (3 equiv.), triethylamine (10 equiv.), and intermediate 1 (1 equiv.) was stirred in dioxane and water (2:1 ratio) at 90 °C until observed by LC/MS the complete consumption of starting material. The solution was diluted with aqueous 1N hydrochloric acid and dichloromethane. The layers were then separated and the aqueous layer was extracted with dichloromethane. The organic substances were combined, dried over magnesium sulfate, filtered, and the solvent was removed in vacuo. Purification produced the desired product.

General procedure C

A mixture of intermediate 2 (this intermediate is described in previously published patent application WO2012/3405 A1; 1 equivalent) and carboxylic acid (1.1 equivalent) in W,W-dimethylformamide was treated with triethylamine (4 equivalents). followed by a solution of 50% ethyl acetate in propylphosphonic anhydride ethyl acetate (T3P, 1.4 equivalents). The reaction was heated to 80 °C for 24 h, after which the reaction was diluted with water and 1N hydrochloric acid solution. The content was extracted with dichloromethane and then ethyl acetate. The combined organic layers were dried over sodium sulfate, filtered, and concentrated in vacuo. Purification produced the desired product.

Pharmaceutically acceptable salts of the invention.

A pharmaceutically acceptable salt may include the inclusion of another molecule, such as an acetate ion, a succinate ion, or another counterion. The counterion can be any organic or inorganic moiety that stabilizes the charge in the precursor compound. Furthermore, a pharmaceutically acceptable salt may have more than one charging atom in its structure. Cases where multiply charged atoms are part of the pharmaceutically acceptable salt may have multiple counterions. Accordingly, a pharmaceutically acceptable salt may have one or more charged atoms and/or one or more counterions.

Pharmaceutically acceptable salts of the claimed compound include those derived from compounds with inorganic acids, organic acids or bases. In some embodiments, the salts can be prepared in situ during the isolation and final purification of the compounds. In other embodiments, the salts can be prepared from the free form of the compound in a separate synthesis step.

Suitable "pharmaceutically acceptable salts" refers to salts prepared from non-toxic pharmaceutically acceptable bases including inorganic bases and organic bases. Salts derived from inorganic bases include those of aluminum, ammonium, calcium, copper, ferric type, ferrous, lithium, magnesium, manganic salts, manganous, potassium, sodium, zinc and the like. Particular embodiments include ammonium, calcium, magnesium, potassium and sodium salts. Salts derived from pharmaceutically acceptable non-toxic organic bases include salts of primary, secondary and tertiary amines, substituted amines including naturally occurring substituted amines, cyclic amines and basic ion exchange resins, such as arginine, betaine, caffeine, choline, N , N.sup.1-dibenzylethylenediamine, diethylamine, 2-diethylaminoethanol, 2-dimethylaminoethanol, ethanolamine, ethylenediamine, N-ethylmorpholine, N-ethylpiperidine, glucamine, glucosamine, histidine, hydrabamine, isopropylamine, lysine, methylglucamine, morpholine, piperazine, piperidine , polyamine resins, procaine, purines, theobromine, triethylamine, trimethylamine tripropylamine, tromethamine and the like.

The salts can be prepared from pharmaceutically acceptable non-toxic acids, including inorganic and organic acids. Such acids include acetic, benzenesulfonic, benzoic, camphorsulfonic, citric, ethanesulfonic, fumaric, gluconic, glutamic, hydrobromic, hydrochloric, isethionic, lactic, maleic, malic, mandelic, methanesulfonic, mucic, nitric, pamoic, pantothenic, phosphoric, succinic, sulfuric, tartaric, p-toluenesulfonic and the like. Particular embodiments include citric, hydrobromic, hydrochloric, maleic, phosphoric, sulfuric and tartaric acids. Other illustrative salts include, but are not limited to, sulfate, citrate, acetate, oxalate, hydrochloric, hydrobromic, iodide, nitrate, bisulfate, phosphate, acid phosphate, isonicotinate, lactate, salicylate, acid citrate, tartrate, oleate, tannate, pantothenate salts. , bitartrate, ascorbate, succinate, maleate, gentisinate, fumarate, gluconate, glucuronate, saccharate, formate, benzoate, glutamate, methanesulfonate, ethanesulfonate, benzenesulfonate, p-toluenesulfonate, and pamoate (i.e., 1,1'-methylene-bis- (2-hydroxy-3-naphthoate)).

The preparation of the pharmaceutically acceptable salts described above and other typical pharmaceutically acceptable salts is described in greater detail in Berg et al., “Pharmaceutical Salts,” J. Pharm. Sci., 1977:66:1-19.

In addition to the compounds described herein, pharmaceutically acceptable salts thereof may also be used in compositions to treat or prevent the disorders identified herein.

Pharmaceutical compositions and methods of administration.

The compounds described herein and pharmaceutically acceptable salts thereof can be formulated as pharmaceutical compositions or “formula ti ons”.

A typical formulation is prepared by mixing a compound, or a pharmaceutically acceptable salt thereof, and a carrier, diluent or excipient. Suitable carriers, diluents and excipients are well known to those skilled in the art and include materials such as carbohydrates, waxes, water-soluble and/or swellable polymers, hydrophilic or hydrophobic materials, gelatin, oils, solvents, water, and the like. The vehicle, diluent or excipient used in each case will depend on the medium and the purpose for which a compound is being formulated. Solvents are generally selected based on solvents recognized by those skilled in the art as safe (GRAS) generally considered safe to be administered to a mammal. In general, safe solvents are non-toxic aqueous solvents, such as water and other non-toxic solvents that are soluble or miscible in water. Suitable aqueous solvents include water, ethanol, propylene glycol, polyethylene glycols (e.g., PEG400, PEG300), etc. and mixtures of these. The formulations may also include other types of excipients such as one or more buffers, stabilizing agents, anti-adherents, surfactants, wetting agents, lubricating agents, emulsifiers, binders, suspending agents, disintegrants, fillers, sorbents, coatings (for example, enteric or slow-release) preservatives, antioxidants, opacifying agents, gliding agents, process adjuvants, colorants, sweeteners, perfume agents, flavoring agents and other additives known to provide elegant presentation of the drug or facilitate the manufacture of the pharmaceutical product (i.e. drug).

The formulations can be prepared using conventional dissolving and mixing procedures. For example, the bulk drug substance (i.e., a compound of claims 1 and 2, pharmaceutically acceptable salts thereof, or a stabilized form of the compounds, such as a complex with a cyclodextrin derivative or other known complexing agent) It is dissolved in a suitable solvent in the presence of one or more of the excipients described above. A compound having the desired degree of purity is optionally mixed with pharmaceutically acceptable diluents, vehicles, excipients or stabilizers, in the form of a lyophilized formulation, ground powder or an aqueous solution. The formulation can be made by mixing at room temperature at the appropriate pH and the desired degree of purity, with physiologically acceptable vehicles. The pH of the formulation depends primarily on the particular use and concentration of the compound, but may vary from about 3 to about 8. When the agent described herein is a solid amorphous dispersion formed by a solvent process, the additives are They can be added directly to the spray drying solution when the mixture is formed, such as when the additive is dissolved or suspended in the solution as a suspension that can then be spray dried. Alternatively, additives can be added after the spray drying process to assist in the formation of the final formulated product.

The compounds of claims 1 and 2 are typically formulated in pharmaceutical dosage forms to provide an easily controllable dosage of the drug and to allow patient compliance with the prescribed regimen. Pharmaceutical formulations of the compounds of claims 1 and 2, or pharmaceutically acceptable salts thereof, can be prepared for various routes and types of administration. Various dosage forms may exist for the same compound, as different medical conditions may warrant different routes of administration.

The amount of active ingredient that can be combined with the carrier material providing a single dosage form will vary depending on the individual treated and the particular mode of administration. For example, a time-release formulation intended for oral administration to humans may contain about 1 to 1,000 mg of active material combined with an appropriate and suitable amount of carrier material that may range from about 5 to about 95% of the compositions. totals (weight: weight). The pharmaceutical composition can be prepared to provide easily quantifiable amounts for administration. For example, an aqueous solution intended for intravenous infusion may contain from about 3 to 500 μg of the active ingredient per milliliter of solution so that the infusion of an adequate volume occurs at a rate of about 30 ml/h. As a general rule, the initial pharmaceutically effective amount of the inhibitor administered will be in the range of about 0.01-100 mg/kg per dose, namely, about 0.1 to 20 mg/kg of patient body weight per day, the typical initial range of compound used being 0.3 to 15 mg/kg/day.

As used herein, the term “therapeutically effective amount” means the amount of pharmaceutical compound or pharmaceutical agent that elicits the biological or medicinal response in a tissue, system, animal or human studied by a researcher, veterinarian, medical doctor. or other clinical professional. The therapeutically or pharmaceutically effective amount of the compound to be administered will be governed by such considerations, and is the minimum amount necessary to ameliorate, cure or treat the disease or disorder or one or more of its symptoms.

The pharmaceutical compositions will be formulated, dosed and administered in a similar manner, that is, quantities, concentrations, schedules, course, vehicles and route of administration, consistent with good medical practice. Factors considered in this context include the particular disorder being treated, the particular mammal being treated, the clinical condition of the individual patient, the cause of the disorder, the site of administration of the agent, the method of administration, the schedule administration and other factors known to medical professionals, such as age, weight, and response of the individual patient.

The term “prophylactically effective amount” refers to an amount effective to prevent or substantially reduce the chances of acquiring a disease or disorder or to reduce the severity of the disease or disorder before acquiring it or to reduce the severity of one or more of its symptoms. before symptoms develop. Roughly, prophylactic measures are divided between primary prophylaxis (to prevent the development of a disease) and secondary prophylaxis (when the disease has already developed and the patient is protected against the worsening of this process).

Acceptable diluents, vehicles, excipients and stabilizers are those that are not toxic to the receptors in the dosages and concentrations used, and include buffers such as phosphate, citrate and other organic acids; antioxidants, including ascorbic acid and methionine; preservatives (such as octadecyldimethylbenzylammonium chloride; hexamethonium chloride; benzalkonium chloride, benzethonium chloride; phenol, butyl or benzyl alcohol; alkylparabens such as methyl or propylparaben; catechol; resorcin; cyclohexanol; 3-pentanol; and m-cresol); proteins, such as serum albumin, gelatin or immunoglobulins; hydrophilic polymers such as polyvinylpyrrolidone; amino acids such as glycine, glutamine, asparagine, histidine, arginine or lysine; monosaccharides, disaccharides and other carbohydrates, including glucose, mannose or dextrins; chelating agents such as EDTA; sugars such as sucrose, mannitol, tretrose or sorbitol; salt-forming counterions such as sodium; metal complexes (for example, Zn protein complexes); and/or nonionic surfactants, such as TWEEN™, PLURONICS™ or polyethylene glycol (PEG). Active pharmaceutical ingredients can also be entrapped in the prepared microcapsules, for example, by coacervate formation techniques or by interfacial polymerization, for example, hydroxymethylcellulose or gelatin microcapsules and poly-(methylmethacrylate) microcapsules, respectively; in colloidal drug delivery systems (e.g., liposomes, albumin microspheres, microemulsions, nanoparticles and nanocapsules) or in macroemulsions. These techniques are described in Remington: The Science and Practice of Pharmacy, 21st edition, University of the Sciences in Philadelphia, Eds., 2005 (hereinafter “Remington”).

“Controlled drug delivery systems” deliver the drug to the body in a precisely controlled manner tailored to the drug and conditions treated. The main objective is to achieve a therapeutic concentration of the drug at the site of action for the desired time. The term “controlled release” is often used to refer to various methods that modify the release of drug from a dosage form. This term includes preparations labeled as “extended release,” “delayed release,” “modified release,” or “sustained release.” In general, controlled release of the agents described herein can be provided by the use of a wide variety of polymeric carriers and controlled release systems including erodible and non-erodible matrices, osmotic control devices, various depot devices, enteric coatings and multiparticle control devices.

“Sustained release preparations” are the most common controlled release applications. Suitable examples of sustained release preparations include semipermeable matrices of solid hydrophobic polymers containing the compound, which matrices are in the form of shaped articles, for example, films or microcapsules. Examples of sustained release matrices include polyesters, hydrogels (e.g., poly(2-hydroxyethyl methacrylate) or poly(vinyl alcohol)), polylactides (US-3,773.9l9), copolymers of L-glutamic acid and ethylene vinyl acetate non-degradable, degradable lactic acid-glycolic acid copolymers, and poly-D-(-)-3-hydroxybutyric acid.

“Immediate release preparations” can also be prepared. The goal of these formulations is to get the drug into the bloodstream and to the site of action as quickly as possible. For example, for rapid dissolution, most tablets are designed to undergo rapid disintegration into granules and subsequent disaggregation into fine particles. This provides a greater specific surface area exposed to the dissolution medium, resulting in a higher dissolution rate.

The agents described herein can be incorporated into an erodible or non-erodible polymeric matrix controlled release device. By erodible matrix, we mean water erodible or water swellable or water soluble in the sense of being erodible or swellable or soluble in pure water or requiring the presence of an acid or base to ionize the polymer matrix sufficiently to cause erosion or dissolution. When it comes into contact with the aqueous environment of use, the erodible polymer matrix absorbs water and forms a gel or swollen aqueous matrix that traps the agent described in the present description. The aqueous swollen matrix gradually erodes, swells, disintegrates or dissolves in the environment of use, thereby controlling the release of a compound described herein to the environment of use. One ingredient of this water-swollen matrix is the swellable, erodible, or water-soluble polymer, which can generally be described as an osmopolymer, hydrogel, or water-swellable polymer. Said polymers can be linear, branched or cross-linked. The polymers can be homopolymers or copolymers. In certain embodiments, they may be synthetic polymers derived from vinyl, acrylate, methacrylate, urethane, ester and oxide monomers. In other embodiments, they may be derived from natural polymers such as polysaccharides (for example, chitin, chitosan, dextran and pullulan; agar gum, gum arabic, karaya gum, locust bean gum, tragacanth gum, carrageenans, ghatti gum, guar gum, gum xanthan and scleroglucan), starches (e.g. dextrin and maltodextrin), hydrophilic colloids (e.g. pectin), phosphatides (e.g. lecithin), alginates (ammonium alginate, sodium, potassium or calcium alginate, propylene glycol alginate) , gelatin, collagen and cellulosic materials. Cellulosic materials are cellulose polymers that have been modified by reacting at least a portion of the hydroxyl groups on the saccharide repeating units with a compound to form an ester-linked or ether-linked substituent. For example, the cellulosic material ethyl cellulose has an ether-linked ethyl substituent attached to the saccharide repeating unit, while the cellulosic material cellulose acetate has an ester-linked acetate substituent. In certain embodiments, the cellulosic materials for the erodible matrix comprise water-soluble and aqueous erodible cellulosic materials that may include, for example, ethyl cellulose (CMC), methyl ethyl cellulose (MEC), carboxymethyl cellulose (CMC), CMEC, hydroxyethyl cellulose (HEC), hydroxypropylcellulose (HPC), cellulose acetate (CA), cellulose propionate (CP), cellulose butyrate (CB), cellulose acetate butyrate (CAB), CAP, CAT, hydroxypropylmethylcellulose (HPMC), HP<m>C<p >, HPMCAS, hydroxypropylmethylcellulose acetate trimethylate (HPMCAT), and ethylhydroxyethylcellulose (EHEC). In certain embodiments, the cellulosic materials comprise various grades of low viscosity (MW less than or equal to 50,000 daltons, for example, Dow Methocel™ series, E5, E15LV, E50LV and K100LY) and high viscosity (MW greater than 50,000 daltons, for example, E4MCR, E10MCR, K4M, K15M and K100M and the Methocel™ K) HPm C series. Other marketed types of HPm C include the Shin Etsu Metolose 90SH series.

Other materials useful as erodible matrix material include, but are not limited to, pullulan, polyvinylpyrrolidone, polyvinyl alcohol, polyvinyl acetate, fatty acid esters of glycerol, polyacrylamide, polyacrylic acid, copolymers of ethacrylic acid or methacrylic acid (EUDRAGIT®, Rohm America, Inc., Piscataway, New Jersey) and other acrylic acid derivatives such as homopolymers and copolymers of butylmethacrylate, methylmethacrylate, ethylmethacrylate, ethylacrylate, (2-dimethylaminoethyl)methacrylate and (trimethylaminoethyl)methacrylate chloride.

Alternatively, the agents of the present invention may be administered or incorporated into a non-erodible matrix device. In these devices, an agent described herein is distributed in an inert matrix. The agent is released by diffusion through the inert matrix. Examples of suitable materials for the inert matrix include insoluble plastics (e.g. methylacrylate-methylmethacrylate copolymers, polyvinyl chloride, polyethylene), hydrophilic polymers (e.g. ethylcellulose, cellulose acetate, cross-linked polyvinylpyrrolidone (also known as crospovidone)) , and fatty compounds (for example, carnauba wax, microcrystalline wax and triglycerides). Such devices are further described in the Remington publication: The Science and Practice of Pharmacy, 20th edition (2000).

As noted above, the agents described herein may also be incorporated into an osmotic control device. Such devices generally include a core containing one or more agents as described herein and a water-permeable, non-soluble, non-erodible coating surrounding the core that controls the flow of water into the core against a aqueous environment of use to cause release of the drug by extrusion of some or all of the cores to the environment of use. In certain embodiments, the coating is polymeric, water permeable and has at least one supply opening. The core of the osmotic device optionally includes an osmotic agent that acts by absorbing water from the surrounding environment through said semipermeable membrane. The osmotic agent contained in the core of this device may be a water-swellable hydrophilic polymer or may be an osmogen, also known as an osmoprotective agent. Pressure is generated within the device, forcing the agent(s) out of the device through an orifice (of a size designed to minimize solute diffusion while preventing the buildup of a hydrostatic pressure head). Non-limiting examples of osmotic control devices are described in United States patent application serial number 09/495,061.

The amount of hydrophilic water-swellable polymers present in the core may vary from about 5 to about 80% by weight (including, for example, from 10 to 50% by weight). Non-limiting examples of core materials include hydrophilic vinyl and acrylic polymers, polysaccharides such as calcium alginate, polyethylene oxide (PEO), polyethylene glycol (PEG), polypropylene glycol (PPG), poly(2-hydroxyethyl methacrylate), poly( acrylic acid), poly(methacrylic acid), polyvinylpyrrolidone (PVP) and cross-linked PVP, poly(vinyl alcohol) (PVA), PVA/PVP copolymers and PVA/PVP copolymers with hydrophobic monomers such as methyl methacrylate, vinyl acetate , and the like, hydrophilic polyurethanes containing large blocks of PEO, croscarmellose sodium, carrageenan, hydroxyethyl cellulose (HEC), hydroxypropyl cellulose (HPC), hydroxypropylmethyl cellulose (HPMC), carboxymethyl cellulose (CMC) and carboxyethyl cellulose (CEC), sodium alginate, polycarbophil , gelatin, xanthan gum and sodium starch glycolate. Other materials include hydrogels comprising interpenetrating networks of polymers that may be formed by addition or condensation polymerization, the components of which may comprise hydrophilic and hydrophobic monomers such as those just mentioned. Hydrophilic water-swellable polymers include, but are not limited to, PEO, PEG, PVP, croscarmellose sodium, HPMC, sodium starch glycolate, polyacrylic acid, and cross-linked versions or mixtures thereof.

The core may also include an osmogen (or osmotic agent). The amount of osmotic agent present in the core may vary from about 2 to about 70% by weight (including, for example, from 10 to 50% by weight). Typical classes of suitable osmogens are water-soluble organic acids, salts and sugars that are capable of imbibing water thereby providing an osmotic pressure gradient across the surrounding coating barrier. Typical useful osmogens include, but are not limited to, magnesium sulfate, magnesium chloride, calcium chloride, sodium chloride, lithium chloride, potassium sulfate, sodium carbonate, sodium sulfite, lithium sulfate, lithium chloride. potassium, sodium sulfate, mannitol, xylitol, urea, sorbitol, inositol, raffinose, sucrose, glucose, fructose, lactose, citric acid, succinic acid, tartaric acid, and mixtures of these. In certain embodiments, the osmogen is glucose, lactose, rose sacchar, mannitol, xylitol, sodium chloride, including combinations of these.

The drug delivery rate is controlled by factors such as the permeability and thickness of the coating, the osmotic pressure of the drug-containing layer, the degree of hydrophilicity of the hydrogel layer, and the specific surface area of the device. Those skilled in the art will appreciate that increasing the thickness of the coating will reduce the release rate, while any of the following will increase the release rate: increasing the permeability of the coating; increase the hydrophilicity of the hydrogel layer; increasing the osmotic pressure of the drug-containing layer; or increase the specific surface area of the device.

In certain embodiments, entrainment of the agent particles described herein in the extrusion fluid during operation of said osmotic device is desirable. For particles to be adequately entrained, the agent drug form is dispersed in the fluid before the particles settle into the tablet core. One means of achieving this is to add a disintegrant which serves to disintegrate the compressed core into its particulate components. Non-limiting examples of standard disintegrants include materials such as sodium starch glycolate (e.g., Explotab™ CLV), microcrystalline cellulose (e.g., Avicel™), silicified microcrystalline cellulose (e.g., ProSoIv™), and croscarmellose sodium (e.g., Ac-Di-Sol™), and other disintegrants known to those skilled in the art. Depending on the specific formulation, some disintegrants give better results than others. Various disintegrants tend to form gels as they swell with water, thereby preventing drug delivery from the device. Non-swelling, non-gelling disintegrants provide faster dispersion of drug particles within the core as water enters the core. In some embodiments, the non-gelling, non-swelling disintegrants are resins, for example, ion exchange resins. In one embodiment, the resin is Amberlite™ IRP 88 (available from Rohm and Haas, Philadelphia, PA). When used, the disintegrant is present in amounts ranging from about 1-25% of the core agent.

Another example of an osmotic device is an osmotic capsule. The capsule shell or part of the capsule shell may be semipermeable. The capsule may be filled with a powder or liquid consisting of an agent described herein, water-imbibing excipients to provide an osmotic potential and/or a water-swellable polymer or, optionally, solubilizing excipients. The capsule core may also be made to have a bilayer or multilayer agent analogous to the bilayer, trilayer or concentric geometries described above.

Another class of osmotic devices useful in this invention comprises coated swellable tablets, for example, as described in EP378404. The coated swelling tablets comprise a tablet core comprising an agent described in the present description and a swelling material, preferably a hydrophilic polymer, coated with a membrane containing holes or pores through which, in the aqueous use environment , the hydrophilic polymer can extrude and extract the agent. Alternatively, the membrane may contain polymeric or low molecular weight water-soluble porosigens. The porosigens dissolve in the aqueous use environment, providing pores through which the hydrophilic polymer and agent can be extruded. Examples of porosigens are water-soluble polymers, such as HPMC, PEG and low molecular weight compounds, such as glycerol, sucrose, glucose and sodium chloride. Additionally, pores can be formed in the coating by drilling holes in the coating with a laser or other mechanical means. In this class of osmotic devices, the membrane material may comprise any film-forming polymer, including polymers that are permeable or impermeable to water, provided that the membrane deposited on the core of the tablet is porous or contains water-soluble porosigens or has a macroscopic hole. for water entry and drug release. Embodiments of this class of sustained release devices may also be multilayer, as described, for example, in EP378404.

When an agent described herein is a liquid or oil, such as a lipid carrier formulation, for example, as described in WO05/011634, the osmotic-type controlled release device may comprise a gel capsule. soft or soft gelatin formed with a wall of composite material and comprising the liquid formulation, where the wall comprises a barrier layer formed on the external surface of the capsule, an expandable layer formed on the barrier layer and a semipermeable layer formed on the layer expandable. A supply opening connects the liquid formulation to the aqueous use environment. Such devices are described, for example, in US6419952, US6342249, US5324280, US4672850, US4627850, US4203440 and US3995631.

As noted above, the agents described herein may be provided as microparticulate materials, generally in a size range of about 10 pm to about 2 mm (including, for example, about 100 pm to 1 mm in diameter). diameter). Such microparticle materials may be packaged, for example, in a capsule, such as a gelatin capsule or a capsule formed from an aqueous soluble polymer, such as HPMCAS, HPMC or starch; be dosed as a heterogeneous solid-liquid suspension or dispersion in a liquid; or they may be formed into a tablet, elongated tablet or pill by compression or other processes known in the art. Such microparticulate materials can be obtained by any known process, such as wet and dry granulation processes, extrusion/spheronization, roller compaction, melt solidification or spray coating seed cores. rization. For example, in wet and dry granulation processes, the agent described in this description and the exceptions Optional elements can be granulated to form multi-particulated materials of the desired size.

The agents can be incorporated into microemulsions, which are generally isotopically transparent and thermodynamically stable dispersions of two immiscible liquids, such as oil and water, stabilized by an interface film of surfactant molecules (Encyclopedia of Pharmaceutical Technology, New York: Marcel Dekker, 1992). , volume 9). For the preparation of microemulsions, surfactant (emulsifier), auxiliary surfactant (auxiliary emulsifier), an oil phase and an aqueous phase are needed. Suitable surfactants include any surfactants useful in the preparation of emulsions, for example, emulsifiers that are typically used in the preparation of creams. The auxiliary surfactant (or “auxiliary emulsifier”) is generally selected from the group of polyglycerol derivatives, glycerol derivatives and fatty alcohols. Preferred emulsifier/auxiliary emulsifier combinations are generally but not necessarily selected from the group consisting of: glyceryl monostearate and polyoxyethylene stearate; polyethylene glycol and ethylene glycol palmitostearate; and caprylic and capric triglycerides and oleyl macrogoglycerides. The aqueous phase includes not only water but also, typically, buffers, glucose, propylene glycol, polyethylene glycols, preferably low molecular weight polyethylene glycols (for example, PEG 300 and PEG 400), and/or glycerol, and the like, while the Oil phase will generally comprise, for example, fatty acid esters, modified vegetable oils, silicone oils, mixtures of monoglycerides, diglycerides and triglycerides, monoesters and diesters of PEG (for example, oleoyl macrogol glycerides), etc.

The compounds described herein can be incorporated into pharmaceutically acceptable nanoparticle, nanosphere and nanocapsule formulations (Delie and Blanco-Prieto, 2005, Molecule 10:65-80). Nanocapsules can generally trap compounds stably and reproducibly. To avoid side effects due to intracellular polymer overload, ultrafine particles (with sizes around 0.1 μm) can be designed using polymers capable of degrading in vivo (e.g., biodegradable polyalkylcyanoacrylate nanoparticles). Such particles are described in the prior art.

Implantable devices coated with a compound of this invention are another embodiment of the present invention. Compounds can also be coated onto implantable medical devices, such as beads or beads, or co-formulated with a polymer or other molecule, to provide a "drug reservoir", thereby allowing the drug to be released over a period of time. longer than the administration of an aqueous solution of the drug. Suitable coatings and general preparation of coated implantable devices are described in US Patents 6,099,562; US-5,886,026; and US-5,304,121. The coatings are typically biocompatible polymeric materials such as a hydrogel polymer, polymethyldisiloxane, polycaprolactone, polyethylene glycol, polylactic acid, ethylene vinyl acetate, and mixtures thereof. Optionally, the coatings may be further coated by a suitable topcoat of fluorosilicone, polysaccharides, polyethylene glycol, phospholipids or combinations thereof to convey controlled release characteristics in the composition.

The formulations include those suitable for the routes of administration detailed in the present description. The formulations may conveniently be presented in unit dosage form and may be prepared by any of the methods well known in the art of pharmacy. The techniques and formulations are generally found in the Remington. Said methods include the step of associating the active ingredient with the vehicle that constitutes one or more auxiliary ingredients. In general, formulations are prepared by bringing the active ingredient into uniform and intimate association with liquid carriers or finely divided solid carriers, or both, and then, if necessary, shaping the product.

The terms "administer", "administering" or "administration" in reference to a compound, composition or formulation of the invention means introducing the compound into the system of the animal in need of treatment. When a compound of the invention is provided in combination with one or more additional active agents, “administration” and its variants are understood to include the simultaneous and/or sequential introduction of the compound and the other active agents.

The compositions described herein may be administered systemically or locally, for example: orally (for example, by capsules, powders, solutions, suspensions, tablets, sublingual tablets and the like), by inhalation (for example, with an aerosol , gas, inhaler, nebulizer or similar), to the ear (for example, using ear drops), topically (for example, using creams, gels, liniments, lotions, ointments, pastes, transdermal patches, etc.), ophthalmically (for example, with eye drops, ophthalmic gels, ophthalmic ointments), rectally (for example, using enemas or suppositories), nasally, buccally, vaginally (for example, using vaginal douches, intrauterine devices, vaginal suppositories, vaginal rings or tablets, etc. .), through an implanted reservoir or similar, or parenterally depending on the severity and type of the disease to be treated. As used herein, the term “parenteral” includes, but is not limited to, subcutaneous, intravenous, intramuscular, intra-articular, intrasynovial, intrasternal, intratrathecal, intrahepatic, intralesional, and intracranial infusion or injection techniques. Preferably, the compositions are administered orally, intraperitoneally or intravenously.

The pharmaceutical compositions described herein may be administered orally in any acceptable oral dosage form including, but not limited to, capsules, tablets, suspensions or aqueous solutions. Liquid dosage forms for oral administration include, but are not limited to, pharmaceutically acceptable emulsions, microemulsions, solutions, suspensions, syrups and elixirs. In addition to the active compounds, liquid dosage forms may contain inert diluents commonly used in the art such as, for example, water or other solvents, solubilizing and emulsifying agents such as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate , benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, dimethylformamide, oils (in particular, cotton, peanut, corn, germ, olive, castor, and sesame oils), glycerol, tetrahydrofurfuryl alcohol, polyethylene glycols and esters of sorbitan fatty acid, and mixtures of these. In addition to inert diluents, oral compositions may also include adjuvants, such as wetting agents, emulsifying and suspending agents, sweeteners, flavors and perfumes.

Solid dosage forms for oral administration include capsules, tablets, pills, powders and granules. In such solid dosage forms, the active compound is mixed with at least one inert pharmaceutically acceptable excipient or vehicle, such as sodium citrate or dicalcium phosphate and/or a) fillers or extenders such as starches, lactose, sucrose, glucose, mannitol, and silicic acid, b) binders such as, for example, carboxymethylcellulose, alginates, gelatin, polyvinylpyrrolidone, sucrose and acacia, c) humectants such as glycerol, d) disintegrating agents such as agar-agar, calcium carbonate, starch potato or tapioca, alginic acid, certain silicates, and sodium carbonate, e) solution retarding agents such as paraffin, f) absorption accelerators such as quaternary ammonium compounds, g) wetting agents such as, for example, cetyl alcohol and glycerol monostearate, h) absorbents such as kaolin and bentonite clay, and i) lubricants such as talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate, and mixtures thereof. The tablets may be uncoated or coated by known techniques, including microencapsulation to mask an unpleasant taste or delay disintegration and adsorption in the gastrointestinal tract and thus provide sustained action over a longer period. For example, a time delay material, such as glyceryl monostearate or glyceryl distearate, may be used alone or with a wax. A water-soluble flavor masking material may be used, such as hydroxypropylmethylcellulose or hydroxypropylcellulose.

Formulations of a compound that are suitable for oral administration may be prepared as discrete units such as tablets, pills, lozenges, throat lozenges, aqueous or oily suspensions, dispersible powders or granules, emulsions, hard or soft capsules, etc. . e.g. gelatin capsules, syrups or elixirs. Formulations of a compound intended for oral use may be prepared according to any method known in the art for the manufacture of pharmaceutical compositions.

Tablets (compressed tablets) may be prepared by compressing in a suitable machine the active ingredient into a free-flowing form such as a powder or granules, optionally mixed with a binder, lubricant, inert diluent, preservative, surfactant or dispersing agent. Molded tablets may be made by molding in a suitable machine a mixture of the powdered active ingredient moistened with an inert liquid diluent.

Formulations for oral use may also be presented as hard gelatin capsules where the active ingredient is mixed with an inert solid diluent, for example, calcium carbonate, calcium phosphate or kaolin, or as soft gelatin capsules where the active ingredient It is mixed with a water-soluble carrier such as polyethylene glycol or an oil medium, for example, peanut oil, liquid paraffin or olive oil.

The active compounds may also be in microencapsulated form with one or more excipients, as mentioned above.

When aqueous suspensions are required for oral use, the active ingredient is combined with emulsifying and suspending agents. If desired, certain sweetening and/or flavoring agents can be added. Syrups and elixirs can be formulated with sweetening agents, for example glycerol, propylene glycol, sorbitol or sucrose. Said formulations may also contain a demulcent, a preservative, flavoring and coloring agents and antioxidant.

Sterile injectable forms of the compositions described herein (e.g., for parenteral administration) may be an aqueous or oleaginous suspension. These suspensions can be formulated according to techniques known in the art using suitable dispersing or wetting agents and suspending agents. The sterile injectable preparation may also be a sterile injectable solution or suspension in a non-toxic parenterally acceptable diluent or solvent, for example, as a solution in 1,3-butanediol. Acceptable vehicles and solvents that can be used include water, Ringer's solution, and isotonic sodium chloride solution. Furthermore, fixed and sterile oils are conventionally used as a solvent or suspending medium. For this purpose, any mild fixed oil can be used, including synthetic monoglycerides or diglycerides. Fatty acids, such as oleic acid and its glyceride derivatives, are useful in the preparation of injectable substances, as are pharmaceutically acceptable natural oils, such as olive oil or castor oil, especially in their polyoxyethylated versions. These oil solutions or suspensions may also contain a long chain alcohol diluent or dispersant, such as carboxymethyl cellulose or similar dispersing agents that are commonly used in the formulation of pharmaceutically acceptable dosage forms including emulsions and suspensions. Other commonly used surfactants, such as Tweens, Spans and other emulsifying agents or bioavailability enhancers that are commonly used in the manufacture of solid, liquid or other pharmaceutically acceptable dosage forms, may also be used for formulation purposes. injectables.

Oil suspensions can be formulated by suspending a compound in a vegetable oil, for example, peanut oil, olive oil, sesame oil or coconut oil or in mineral oil such as liquid paraffin. Oily suspensions may contain a thickening agent, for example beeswax, hard paraffin or cetyl alcohol. Sweetening agents, such as those described above, and flavoring agents may be added to provide a palatable oral preparation. These compositions can be preserved by the addition of an antioxidant, such as butylated hydroxyanisole or alpha-tocopherol.

Aqueous suspensions of a compound contain the active materials in a mixture with excipients suitable for the manufacture of aqueous suspensions. Such excipients include a suspending agent, such as sodium carboxymethylcellulose, croscarmellose, povidone, methylcellulose, hydroxypropylmethylcellulose, sodium alginate, polyvinylpyrrolidone, gum tragacanth and gum acacia, and dispersing or wetting agents such as a natural phosphatide (for example, lecithin ), a condensation product of an alkylene oxide with a fatty acid (e.g., polyoxyethylene stearate), a condensation product of ethylene oxide with a long-chain aliphatic alcohol (e.g., heptadecaethyleneoxycetanol), a condensation product of ethylene oxide with a partial ester derived from a fatty acid and a hexitol anhydride (for example, polyoxyethylene sorbitan monooleate). The aqueous suspension may also contain one or more preservatives such as ethyl or n-propyl p-hydroxybenzoate, one or more coloring agents, one or more flavoring agents and one or more sweetening agents, such as sucrose or saccharin.

Injectable formulations can be sterilized, for example, by filtration through a bacterial retention filter, or by incorporating sterilizing agents in the form of sterile solid compositions that can be dissolved or dispersed in sterile water or other aqueous injectable medium before use. .

In order to prolong the effect of a compound described herein, it is often desirable to slow the absorption of the compound from subcutaneous or intramuscular injection. This can be achieved by using a liquid suspension of crystalline or amorphous material with poor solubility in water. The absorption rate of the compound depends, then, on its dissolution rate which, in turn, may depend on the crystal size and crystalline form. Alternatively, delayed absorption of a parenterally administered form of compound is achieved by dissolving or suspending the compound in an oil vehicle. Long-acting injectable forms are made by forming microencapsulated matrices of the compound in biodegradable polymers, such as polylactide-polyglycolide. Depending on the ratio of the compound to the polymer and the nature of the specific polymer used, the release rate of the compound can be controlled. Examples of other biodegradable polymers include poly(orthoesters) and poly(anhydrides). Long-acting injectable formulations are also prepared by trapping the compound in liposomes or microemulsions that are compatible with body tissues.

Injectable solutions or microemulsions can be introduced into a patient's bloodstream by local bolus injection. Alternatively, it may be advantageous to administer the solution or microemulsion in such a way that a constant circulating concentration of the compound in question is maintained. To maintain that constant concentration, a continuous intravenous delivery device can be used. An example of this device is the Deltec CADD-PLUS™ model 5400 intravenous pump.

Compositions for rectal or vaginal administration are preferably suppositories which can be prepared by mixing the compounds described herein with suitable non-irritating excipients or vehicles such as cocoa butter, beeswax, polyethylene glycol or a suppository wax that is solid at temperature. ambient but liquid at body temperature and therefore melt in the rectum or vaginal cavity and release the active compound. Other formulations suitable for vaginal administration may be presented as pessaries, tampons, creams, gels, pastes, foams or sprays.

The pharmaceutical compositions described herein can also be administered topically, especially when the treatment target includes areas or organs easily accessible by topical application, including diseases of the eye, ear, skin or lower intestinal tract. Suitable topical formulations are easily prepared for each of these areas or organs.

Dosage forms for topical or transdermal administration of a compound described herein include ointments, pastes, creams, lotions, gels, powders, solutions, sprays, inhalants or patches. The active component is mixed under sterile conditions with a pharmaceutically acceptable carrier and any preservatives or buffers that may be necessary. Also contemplated within the scope of the invention is an ophthalmic formulation, ear drops and eye drops. Additionally, the present invention contemplates the use of transdermal patches, which have the added advantage of providing a controlled delivery of a compound to the body. These dosage forms can be prepared by dissolving or dispensing the compound in the appropriate medium. Absorption enhancers can also be used to increase the flow of the compound throughout the skin. The rate can be controlled by providing a rate controlling membrane or by dispersing the compound in a polymer matrix or gel. Topical application to the lower intestinal tract may be in a rectal suppository formulation (see above) or in a suitable enema formulation. Transdermal patches can also be used topically.

For topical applications, the pharmaceutical compositions can be formulated in a suitable ointment containing the active component suspended or dissolved in one or more vehicles. Vehicles for topical administration of the compounds of this invention include, but are not limited to, mineral oil, liquid petroleum jelly, white petroleum jelly, propylene glycol, polyoxyethylene, polyoxypropylene compound, emulsifying wax and water. Alternatively, the pharmaceutical compositions may be formulated in a suitable lotion or cream containing the active components suspended or dissolved in one or more pharmaceutically acceptable carriers. Suitable carriers include, but are not limited to, mineral oil, sorbitan monostearate, polysorbate 60, cetyl esters wax, cetearyl alcohol, 2-octyldodecanol, benzyl alcohol and water.

For ophthalmic use, the pharmaceutical compositions may be formulated as micronized suspensions in isotonic, pH-adjusted sterile saline, or, preferably, as solutions in isotonic, pH-adjusted sterile saline, either with or without a preservative, such as sodium chloride. benzylalkonium. Alternatively, for ophthalmic uses, the pharmaceutical compositions may be formulated in an ointment such as petroleum jelly. For the treatment of the eye or other external tissues, for example, the mouth and skin, the formulations may be applied as a topical ointment or cream containing the active ingredient(s) in an amount of, for example, 0.075 to 20 wt%. /p. When formulated into an ointment, the active ingredients may be employed with an oil-based, paraffinic, or water-miscible ointment base.

Alternatively, the active ingredients may be formulated in a cream with an oil in an oil/water cream base. If desired, the aqueous phase of the cream base may include a polyhydric alcohol, that is, an alcohol having two or more hydroxyl groups, such as propylene glycol, butane 1,3-diol, mannitol, sorbitol, glycerol and polyethylene glycol ( including PEG 400) and mixtures of these. Topical formulations may desirably include a compound that enhances the absorption or penetration of the active ingredient through the skin or other affected areas. Examples of such dermal penetration enhancers include dimethyl sulfoxide and related analogs.

The oil phase of emulsions prepared using a compound may consist of known ingredients in a known manner. While the phase may simply comprise an emulsifier (also known as an emulsifier), it preferably comprises a mixture of at least one emulsifier with a fat or an oil or with a fat and an oil. A hydrophilic emulsifier may be included along with a lipophilic emulsifier that acts as a stabilizer. In some embodiments, the emulsifier includes both an oil and a fat. Together, the emulsifier(s) with or without stabilizer(s) constitute the so-called emulsifying wax and the wax together with the oil and fat constitute the so-called emulsifying ointment that forms the dispersed oil phase of the cream formulations. Emulsifiers and emulsion stabilizers suitable for use in formulating a compound include Tween™-60, Span™-80, cetostearyl alcohol, benzyl alcohol, myristyl alcohol, glyceryl monostearate and sodium lauryl sulfate.

The pharmaceutical compositions may further be administered by nasal spray or inhalation. Such compositions are prepared according to techniques well known in the art of pharmaceutical formulation and may be prepared as solutions in saline, using benzyl alcohol or other suitable preservatives, absorption promoters to improve bioavailability, fluorocarbons and/or other conventional solubilizing or dispersing agents. . Formulations suitable for intrapulmonary or nasal administration have a particle size, for example, in the range of 0.1 to 500 micrometers (including particles in a range between 0.1 and 500 micrometers in increments of 0.5, 1, 30, 35 micrometers, etc.) that are administered by rapid inhalation through the nasal passage or by inhalation through the mouth to reach the alveolar sacs.

The pharmaceutical composition (or formulation) for use can be packaged in a variety of ways depending on the method used to administer the drug. Generally, an article for distribution includes a container having the pharmaceutical formulation deposited therein in a suitable form. Suitable containers are well known to those skilled in the art and include materials such as bottles (plastic and glass), sachets, blisters, plastic bags, metal cylinders and the like. The container may also include an anti-tamper assembly to prevent intrusive access to the contents of the container. In addition, the container has a label placed on it that describes the contents of the container. The label may also include appropriate warnings.

The formulations may be packaged in unit dose or multi-dose containers, for example, sealed ampoules and vials, and may be stored in a freeze-dried (lyophilized) condition, requiring only the addition of the sterile liquid carrier, for example water, for a injection immediately before use. Extemporaneous injection solutions and suspensions are prepared from sterile powders, granules and tablets of the type described above. Preferred unitary pharmaceutical formulations are those that contain a daily dose or unit less than the daily dose, as mentioned above in the present description, or a suitable fraction thereof, of the active ingredient.

In another aspect, a compound or a pharmaceutically acceptable salt thereof may be formulated in a veterinary composition comprising a veterinary carrier. Veterinary carriers are materials useful for the purpose of administration of the composition and may be solid, liquid or gaseous materials that are otherwise inert and compatible with the active ingredient. These veterinary compositions can be administered parenterally, orally or by any other desired route.

Therapeutic methods

The invention also relates to the compounds of claims 1 and 2 for use in the treatment of certain disorders, alone or in combination, or their pharmaceutically acceptable salts or pharmaceutical compositions comprising them.

The present description relates to soluble guanylate cyclase (sGC) stimulators, pharmaceutical formulations thereof and their use, alone or in combination with one or more additional agents, to treat and/or prevent various diseases, where an increase may be desirable. in the concentration of NO or an increase in the concentration of cGMP. Diseases that can be treated include, but are not limited to, pulmonary hypertension, high blood pressure, heart failure, atherosclerosis, inflammation, thrombosis, fibrosis and kidney failure, cirrhosis of the liver, erectile dysfunction, female sex disorders, blood-related disorders. diabetes, eye disorders and other related cardiovascular disorders.

The increased concentration of cGMP produces vasodilation, inhibition of platelet aggregation and adhesion, antihypertensive effects, anti-remodeling effects, anti-apoptotic effects, anti-inflammatory effects, and neuronal signaling effects. Therefore, sGC stimulators can be used to treat and/or prevent a range of diseases and disorders, including, but not limited to, peripheral, pulmonary, hepatic, liver, cardiac or cerebrovascular/endothelial disorders or conditions, a urogenitalgynecological or sexual disorder or condition, a thromboembolic disease, an ischemic disease, a fibrotic disorder, a topical or dermal disorder, a pulmonary or respiratory disorder, a renal or hepatic disorder, a metabolic disorder, atherosclerosis, or a lipid-related disorder .

The compounds described herein are sGC stimulators that may be useful in the prevention and/or treatment of diseases and disorders that are characterized by having an undesirably reduced bioavailability of and/or sensitivity to NO(s), such as those associated with oxidative stress or nitrosative stress conditions.

Throughout this description, the terms “hypertension,” “high blood pressure,” or “high blood pressure (HBP)” are used interchangeably and refer to an extremely common and highly preventable chronic condition in which blood pressure (blood pressure - BP) in the arteries is higher than normal. If not properly controlled, it is a significant risk factor for several serious cardiovascular and kidney conditions. Hypertension can be a primary disease, called “essential hypertension” or “idiopathic hypertension,” or it can be caused by other diseases, in which case it is classified as “secondary hypertension.” Essential hypertension accounts for 90-95% of all cases.

As used herein, the term “resistant hypertension” refers to hypertension that remains above the target blood pressure (usually less than 140/90 mmHg, although a lower target of less than 130/80 mmHg is recommended. , for patients with comorbid diabetes or kidney disease), despite the simultaneous use of three antihypertensive agents belonging to different antihypertensive drug classes. Additionally, people who require four or more drugs to control their blood pressure are considered to have resistant hypertension. Hypertension is an extremely common condition that is comorbid with diabetes, affecting ~20-60% of patients with diabetes, depending on obesity, ethnicity, and age. This type of hypertension is referred to herein as “diabetic hypertension.” In type 2 diabetes, hypertension is usually present as part of the metabolic syndrome of insulin resistance, also including central obesity and dyslipidemia. In type 1 diabetes, hypertension may signal the onset of diabetic nephropathy.

“Pulmonary hypertension (PH), as used herein, is a disease characterized by sustained elevations in blood pressure in the pulmonary vasculature (pulmonary artery, pulmonary vein, and pulmonary capillaries), resulting in right cardiac hypertrophy, leading to ultimately to right heart failure and death. Common symptoms of PH include difficulty breathing, dizziness, and fainting, which are exacerbated by exertion. Without treatment, the average life expectancy after diagnosis is 2.8 years. PH exists in many different forms, which are classified according to their etiology. Categories include pulmonary arterial hypertension (PAH), PH with left heart disease, PH associated with lung diseases and/or hypoxemia, PH due to chronic thrombotic and/or embolic disease, and miscellaneous PH. PAH is rare in the general population, but prevalence increases in association with certain common conditions such as HIV infection, scleroderma, and sickle cell disease. Other forms of PH are generally more common than PAH and, for example, the association of PH with chronic obstructive pulmonary disease (COPD) is of particular interest. The current treatment of pulmonary hypertension depends on the stage and mechanism of the disease.

As used herein, “heart failure” is a progressive disorder of left ventricular (LV) myocardial remodeling culminating in a complex clinical syndrome in which impaired cardiac function and circulatory congestion are the defining features. and results in insufficient supply of blood and nutrients to body tissues. The condition occurs when the heart is damaged or overworked and cannot pump all the blood returned to it from its systemic circulation. As less blood is pumped, blood returning to the heart pools and fluid builds up in other parts of the body. Heart failure also affects the kidneys' ability to dispose of sodium and water, further complicating fluid retention. Heart failure is characterized by autonomic dysfunction, neurohormonal activation, and cytokine overproduction, which contribute to progressive circulatory failure. Symptoms of heart failure include: dyspnea (difficulty breathing) when exercising and waking up at night due to sudden onset of breathing difficulties, both of which are indicative of pulmonary edema; general fatigue or weakness, edema of the feet, ankles and legs, rapid weight gain, chronic cough, including cough with production of mucus or blood. Depending on its clinical presentation, heart failure is classified as de novo, transient or chronic heart failure. Acute heart failure, that is, the rapid or gradual onset of symptoms requiring urgent therapy, may develop de novo or as a result of decompensation of heart failure. Diabetes is a common comorbidity in patients with heart failure and is associated with a worse prognosis as well as the possibility that the effectiveness of treatments will be affected. Other important comorbidities include systemic hypertension, chronic airflow obstruction, sleep apnea, cognitive dysfunction, anemia, chronic kidney disease, and chronic arthritis. Chronic left heart failure is often associated with the development of pulmonary hypertension. The frequency of certain comorbidities varies depending on sex: among women, hypertension and thyroid disease are more common, while men more commonly suffer from chronic obstructive pulmonary disease (COPD), peripheral vascular disease, coronary artery disease and kidney failure. Depression is a common comorbidity of heart failure, and the two conditions can often complicate each other. Cachexia has long been recognized as a serious and common complication of heart failure, affecting up to 15% of all heart failure patients and associated with a poor prognosis. Cardiac cachexia is defined as the involuntary, non-edematous loss of at least 6% of body weight over a six-month period.

The term “sleep apnea” refers to the most common sleep-disrupting breathing disorder. It is a condition characterized by intermittent, cyclical reductions or complete interruptions in airflow, which may or may not involve upper airway obstruction. There are three types of sleep apnea: obstructive sleep apnea, the most common form, central sleep apnea, and mixed sleep apnea.

“Central sleep apnea (CSA)” is caused by a disruption of the brain's normal signal to breathe, rather than a physical blockage of the airways. Lack of respiratory effort results in an increase in carbon dioxide in the blood, which can wake up the patient. ACS is rare in the general population, but is a relatively common manifestation in patients with systolic heart failure.

As used herein, the term “metabolic syndrome,” “insulin resistance syndrome,” or “syndrome X” refers to a group or cluster of metabolic conditions (abdominal obesity, elevated fasting glucose, and (elevated blood pressure (HBP)), which occur more frequently together than in isolation and which jointly promote the development of type 2 diabetes and cardiovascular disease. Metabolic syndrome is characterized by a specific lipid profile of increased triglycerides, reduced levels of high-density lipoprotein-cholesterol (HDL-cholesterol) and, in some cases, moderately high levels of low-density lipoprotein-cholesterol (LDL-cholesterol), as well as accelerated progression of “atherosclerotic disease.” ” due to the pressure of component risk factors. There are several types of dyslipidemias: “Hypercholesterolemia” refers to elevated cholesterol levels. Familial hypercholesterolemia is a specific form of hypercholesterolemia due to a defect in chromosome 19 (19p13, 1-13,3). “Hyperglyceridemia” refers to elevated levels of glycerides (for example, “hypertriglyceridemia” implies elevated levels of triglycerides). “Hyperlipoproteinemia” refers to elevated levels of lipoproteins (usually LDL, unless otherwise noted).

As used herein, the term “peripheral vascular disease (PVD)”, also commonly referred to as “peripheral arterial disease (PAD)” or “peripheral arterial occlusive disease (PAOD)” refers to the obstruction of large arteries that form part of the coronary vasculature, aortic arch or brain. PVD may be due to atherosclerosis, inflammatory processes leading to stenosis, embolism, or thrombus formation. It produces acute or chronic “ischemia (lack of blood supply)”. PVD is often a term used to refer to atherosclerotic obstructions present in the lower extremity. PVD also includes a subset of diseases classified as microvascular diseases that are due to episodic narrowing of the arteries (for example, “Raynaud's phenomenon”) or widening of the arteries (erythromelalgia), that is, vascular spasms.

The term “thrombosis” refers to the formation of a blood clot (“thrombus”) within a blood vessel, obstructing the flow of blood through the circulatory system. When a blood vessel is injured, the body uses platelets (thrombocytes) and fibrin to form a blood clot to prevent blood loss. Alternatively, even if a blood vessel is not injured, blood clots can form in the body if the right conditions are present. If clotting is too pronounced and the clot breaks free, the displaced clot is then known as an “embolus.” The term “thromboembolism” refers to the combination of thrombosis and its main complication, “embolism.” When a thrombus occupies more than 75% of the specific surface area of an artery's lumen, blood flow to the supplied tissue is reduced enough to cause symptoms due to reduced oxygen content (hypoxia) and the accumulation of metabolic products such as lactic acid (“gout”). An obstruction of more than 90% can lead to anoxia, a complete lack of oxygen, and “infarction,” a mode of cell death.

An “embolism” (plural, embolisms) is the case in which an embolus (a separate intravascular mass capable of obstructing arterial capillary beds at a site far from its origin) becomes lodged in a narrow capillary vessel of an arterial bed, resulting in causing an obstruction (vascular occlusion) in a distant part of the body. This should not be confused with a thrombus causing obstruction at the site of origin.

A “stroke” or cerebrovascular accident (CVA) is the rapid loss of brain function or functions due to an alteration in the blood supply to the brain. This may be due to “ischemia” (lack of blood flow) caused by obstruction (thrombosis, arterial embolism), or hemorrhage (leakage of blood). As a result, the affected area of the brain cannot function, which could result in an inability to move one or more limbs on one side of the body, inability to speak or understand speech, or inability to see one side of the visual field. Risk factors for stroke include age, hypertension, a previous stroke or transient ischemic attack (TIA), diabetes, high cholesterol, cigarette smoking, and atrial fibrillation. High blood pressure is the most important modifiable risk factor for stroke. An “ischemic stroke” is occasionally treated in a hospital with thrombolysis (also known as a “thrombus breaker”), and some hemorrhagic strokes benefit from neurosurgery. Prevention of relapses may involve the administration of antiplatelet drugs, such as aspirin and dipyridamole, control and reduction of hypertension, and the use of statins. Selected patients may benefit from carotid endarterectomy and the use of anticoagulants.

“Ischemia” is a restriction in the blood supply to tissues, causing an insufficiency of oxygen and glucose necessary for cellular metabolism (to keep the tissue alive). Ischemia is usually caused by problems with the blood vessels, with resulting tissue damage or dysfunction. It also means local anemia in a given part of the body which is sometimes due to congestion (such as vasoconstriction, thrombosis or embolism).

According to the Diagnostic and Statistical Manual of the American Psychiatric Association of Mental Disorders, Fourth Edition (DSM-IV), the term “sexual dysfunction” encompasses a series of conditions “characterized by alterations in sexual desire and psychophysiological changes associated with the sexual response cycle”; Although problems of this type are common, sexual dysfunction is considered to exist only when the problems cause discomfort to the patient. Sexual dysfunction can be of physical or psychological origin. It may exist as a primary condition, generally hormonal, although it is most often secondary to other medical conditions or pharmacological therapy for such conditions. All types of sexual dysfunction can be further classified as long-term, acquired, situational, or generalized (or combinations of these).

The DSM-IV-TR specifies five major categories of “female sexual dysfunction”: disorders of sexual desire/interest; “sexual desire disorders (including genital, subjective and combined)”; orgasmic disorders; dyspareunia and vaginismus; and persistent sexual desire disorder.

“Female hypoactive sexual desire disorder (HSDD)” is defined as a persistent or recurrent inability to achieve or maintain sufficient levels of sexual arousal, causing suffering to the person. The term “HSDD” encompasses both a lack of subjective feelings of arousal (i.e., subjective sexual desire disorder) and a lack of somatic responses such as lubrication and dilation (i.e., genital/sexual sexual desire disorder). HSDD can be strictly psychological in origin, although it is generally caused or complicated by medical or physiological factors. Hypoestrogenism is the most common physiological state associated with HSDD, leading to urogenital atrophy and decreased vaginal lubrication.

As used herein, “erectile dysfunction (ED)” is a male sexual dysfunction, which is characterized by the inability to develop or maintain a penile erection during sexual intercourse. A penile erection is the hydraulic effect of blood entering and retaining it in the spongy bodies inside the penis. The process often begins as a result of sexual arousal, when signals are transmitted from the brain to the nerves in the penis. Erectile dysfunction is indicated when an erection is difficult to produce. The most important root causes are cardiovascular disease and diabetes, neurological problems (for example, trauma from prostatectomy surgery), hormonal insufficiency (hypogonadism), and adverse drug effects.

As used herein, the term “bronchoconstriction” is used to define the constriction of the airways in the lungs due to tension of the surrounding smooth muscle, with subsequent coughing, labored breathing, and shortness of breath. The condition has a number of causes, the most common being asthma. Exercises and allergies can trigger symptoms in an otherwise asymptomatic individual. Other conditions such as chronic obstructive pulmonary disease (COPD) can also present with bronchoconstriction.

Specific disease disorders that can be treated and/or prevented by administering an sGC stimulator of the invention include, but are not limited to: hypertension (e.g., diabetic hypertension, arterial hypertension, pulmonary hypertension, resistant hypertension, peripheral arterial disease, etc.), heart failure (e.g. left ventricular diastolic dysfunction (LVDD) and left ventricular systolic dysfunction (LVSD), sleep apnea associated with heart failure), sclerotic disease (e.g. atherosclerosis), thromboembolic disorders (e.g. chronic thromboembolic pulmonary hypertension, thrombosis, stroke, embolism, pulmonary embolism), Alzheimer's disease, kidney diseases (e.g., renal fibrosis, ischemic kidney disease, renal failure, renal insufficiency, chronic kidney disease), liver disease (e.g., fibrosis liver disease or cirrhosis), respiratory disease (for example, pulmonary fibrosis, asthma, chronic obstructive pulmonary disease, interstitial lung disease), sexual disorders (for example, erectile dysfunction, male or female sexual dysfunction, vaginal atrophy), sickle cell anemia, neuroinflammatory diseases or associated disorders and diseases (for example, lipid-related disorders).

Another aspect of the invention is the compounds of claims 1 and 2, as well as pharmaceutically acceptable salts thereof, for use in the treatment of diseases, conditions and health disorders selected from:

(1) peripheral, pulmonary, hepatic, renal, cardiac, or cerebrovascular/endothelial disorders/conditions otherwise related to circulation:

• disorders related to high blood pressure and reduced coronary blood flow, such as acute and chronic elevated coronary blood pressure, arterial hypertension and vascular disorder due to complications of the heart and kidney (for example, heart disease, stroke, cerebral ischemia, renal failure ); resistant hypertension, diabetic hypertension, congestive heart failure; diastolic or systolic dysfunction; coronary insufficiency; arrhythmias; reduction of ventricular preload; cardiac hypertrophy; heart/cardiorenal failure syndrome; portal hypertension; endothelial dysfunction or injury;

• and thromboembolic disorders and ischemia such as myocardial infarction, stroke, transient ischemic attacks (TIA); obstructive thromboanginitis; stable or unstable angina pectoris; coronary spasms, variant angina, Prinzmetal's angina; prevention of restenosis after thrombolysis therapies; thrombogenic disorders;

• Alzheimer disease; Parkinson's disease; dementia; vascular cognitive impairment; cerebral vasospasms; head trauma;

• peripheral arterial disease, peripheral arterial occlusive disease; peripheral vascular disease; hypertonia; Raynaud's syndrome or phenomenon, critical limb ischemia, vasculitis; peripheral embolism; intermittent claudication; vaso-occlusive crises; Duchenne and Becker muscular dystrophies; microcirculation abnormalities; control of leaks or vascular permeability;

• shock; septicemia; cardiogenic shock; control of leukocyte activation; inhibition or modulation of platelet aggregation;

• pulmonary/respiratory conditions, such as pulmonary hypertension, pulmonary arterial hypertension and associated pulmonary vascular remodeling (eg, localized thrombosis and right cardiac hypertrophy); pulmonary hypertonia; primary pulmonary hypertension, secondary pulmonary hypertension, familial pulmonary hypertension, sporadic pulmonary hypertension, precapillary pulmonary hypertension, idiopathic pulmonary hypertension, thrombotic pulmonary arteriopathy, plexogenic pulmonary arteriopathy; cystic fibrosis; bronchoconstriction or pulmonary bronchoconstriction; acute respiratory distress syndrome; pulmonary fibrosis, lung transplant;

• pulmonary hypertension associated with or related to: left ventricular dysfunction, hypoxemia, hypertension of WHO groups I, II, III, IV and V, mitral valve disease, constrictive pericarditis, aortic stenosis, cardiomyopathy, mediastinal fibrosis, pulmonary fibrosis , anomalous pulmonary venous drainage, pulmonary veno-occlusive disease, pulmonary vasculitis, collagen vascular disease, congenital heart disease, pulmonary venous hypertension, interstitial lung disease, sleep disordered breathing, sleep apnea, alveolar hypoventilation disorders, chronic exposure to high altitude, disease pulmonary disease in neonates, alveolar capillary dysplasia, sickle cell anemia, other coagulation disorders, chronic thromboembolism, pulmonary embolism (due to tumor, parasites or foreign material), connective tissue disease, lupus, schistosomiasis, sarcoiditis, chronic obstructive pulmonary disease, asthma, emphysema, chronic bronchitis, pulmonary capillary hemangiomatosis; histiocytosis

• atherosclerotic diseases or conditions, such as atherosclerosis (for example, associated with endothelial injury, platelet and monocyte adhesion and aggregation, smooth muscle proliferation and migration); restenosis (for example, developed after thrombolysis therapies, percutaneous transluminal angioplasties (PTA), percutaneous transluminal coronary angioplasties (PTCA), and bypasses); inflammation;

• cardiovascular disease associated with metabolic syndrome (eg, obesity, dyslipidemia, diabetes, high blood pressure); lipid-related disorders, such as dyslipidemia, hypercholesterolemia, hypertriglyceridemia, sitosterolemia, fatty liver disease, and hepatitis; preeclampsia; progression of polycystic kidney disease; subcutaneous fat; obesity;

• cirrhosis of the liver, associated with chronic liver disease, liver fibrosis, activation of hepatic stellate cells, accumulation of hepatic fibrous collagen and total collagen; liver disease of necroinflammatory and/or immunological origin; disorders of the andurogenital system, such as renal fibrosis and renal failure that are due to chronic renal disease or failure (for example, due to tissue accumulation/deposition and injury, progressive sclerosis, glomerulonephritis); prostatic hypertrophy systemic sclerosis; cardiac interstitial fibrosis; cardiac remodeling and fibrosis; cardiac hypertrophy;

(2) ischemia or reperfusion injury; ischemia/reperfusion associated with organ transplant, lung transplant, lung transplant, heart transplant; preservation of blood substituents in trauma patients;

(3) sexual, gynecological and urological disorders: erectile dysfunction; impotence; premature ejaculation; female sexual dysfunction (eg, female sexual desire dysfunction, hypoactive sexual desire disorder), vaginal atrophy, dyspaneuria, atrophic vaginitis; benign prostatic hyperplasia (BPH) or benign prostatic hypertrophy or enlargement, bladder outlet obstruction; painful bladder syndrome (BPS), interstitial cystitis (IC), overactive bladder syndrome, and neurogenic bladder incontinence; diabetic nephropathy;

(4) ocular diseases or disorders: glaucoma, retinopathy, diabetic retinopathy, blepharitis, dry eye syndrome, Sjogren's syndrome;

(5) hearing diseases or disorders: hearing deficiency, partial or total hearing loss; partial or total deafness; tinnitus; noise-induced hearing loss;

(6) topical or skin disorders or conditions: dermal fibrosis, scleroderma, skin fibrosis;

(7) wound healing: for example, in diabetics; improvement of microvascular perfusion (for example, after injury, to counteract the inflammatory response in perioperative care), anal fissures, diabetic ulcers; and

(8) other diseases or conditions: cancer metastasis, osteoporosis, gastroparesis; functional dyspepsia; diabetic complications, diseases associated with endothelial dysfunction and neurological disorders associated with decreased nitric oxide production.

In other embodiments of the invention, the compounds of claims 1 and 2 as well as pharmaceutically acceptable salts thereof, are for use in the prevention and/or treatment of the following types of diseases, conditions and disorders that can be improved with stimulation of sGC:

hypertension, resistant hypertension, diabetic hypertension, pulmonary hypertension (PH), pulmonary arterial hypertension, PH associated with COPD, chronic airflow obstruction, asthma or pulmonary fibrosis, thrombosis, embolism, thromboembolic disorders, Alzheimer's disease, atherosclerosis, cardiac hypertrophy right, heart failure, diastolic dysfunction, systolic dysfunction, sleep apnea associated with heart failure, liver cirrhosis, renal fibrosis, renal failure due to kidney disease or failure, metabolic disorder, dyslipidemia, hypercholesterolemia, hypertriglyceridemia, sitosterolemia, fatty liver disease, hepatitis, erectile dysfunction, female sexual dysfunction, female sexual desire dysfunction and vaginal atrophy.

In other embodiments the disease, condition or health disorder is selected from a peripheral, pulmonary, hepatic, kidney, cardiac or cerebrovascular/endothelial disorder or condition, or a disease otherwise related to the circulation selected from: coronary blood pressure acute and chronic elevated, arterial hypertension and vascular alteration due to heart and kidney complications, heart disease, stroke, cerebral ischemia, renal failure; resistant hypertension, diabetic hypertension, congestive heart failure; diastolic or systolic dysfunction; coronary insufficiency; arrhythmias; reduction of ventricular preload; cardiac hypertrophy; heart/cardiorenal failure syndrome; portal hypertension; endothelial dysfunction or injury; myocardial infarction; stroke or transient ischemic attacks (TIA); obstructive thromboanginitis; stable or unstable angina pectoris; coronary spasms, variant angina, Prinzmetal's angina; restenosis due to thrombolysis therapies and thrombogenic disorders.

In other embodiments, the disease, condition or health disorder is selected from a peripheral vascular/endothelial disorder or condition or a disease otherwise related to the circulation selected from: peripheral arterial disease, peripheral arterial occlusive disease; peripheral vascular disease; hypertonias; Raynaud's syndrome or phenomenon; critical limb ischemia; vasculitis; peripheral embolism; intermittent claudication; vaso-occlusive crises; Duchenne and Becker muscular dystrophies; microcirculation abnormalities; and vascular leak or permeability problems.

In other embodiments, the disease, condition or health disorder is a pulmonary disorder or condition or a disease otherwise related to the circulation selected from: pulmonary hypertension; pulmonary arterial hypertension and associated pulmonary vascular remodeling; localized thrombosis; right cardiac hypertrophy; pulmonary hypertonia; primary pulmonary hypertension, secondary pulmonary hypertension, familial pulmonary hypertension, sporadic pulmonary hypertension, precapillary pulmonary hypertension, idiopathic pulmonary hypertension, thrombotic pulmonary arteriopathy, plexogenic pulmonary arteriopathy; cystic fibrosis; bronchoconstriction or pulmonary bronchoconstriction; acute respiratory distress syndrome; pulmonary fibrosis and lung transplantation. In some of these embodiments, pulmonary hypertension is pulmonary hypertension associated with or related to left ventricular dysfunction, hypoxemia, WHO group I, II, III, IV and V hypertension, mitral valve disease, constrictive pericarditis, stenosis. aortic, cardiomyopathy, mediastinal fibrosis, pulmonary fibrosis, anomalous pulmonary venous drainage, pulmonary veno-occlusive disease, pulmonary vasculitis, collagen vascular disease, congenital heart disease, pulmonary venous hypertension, interstitial lung disease, sleep disordered breathing, sleep apnea, hypoventilation disorders alveolar, chronic exposure to high altitude, pulmonary disease in neonates, alveolar capillary dysplasia, sickle cell anemia, coagulation disorders, chronic thromboembolism; pulmonary embolism, due to tumors, parasites or foreign material; connective tissue disease, lupus, schistosomiasis, sarcoidosis, chronic obstructive pulmonary disease, asthma, emphysema, chronic bronchitis and pulmonary capillary hemangiomatosis; histiocytosis lymphangiomatosis and compressed pulmonary vessels due to adenopathy, tumor or fibrosing mediastinitis.

In other embodiments, the health condition or disorder is a vascular or endothelial disorder or condition or a disease otherwise related to the circulation selected from: atherosclerotic diseases; atherosclerosis, atherosclerosis associated with endothelial injury, atherosclerosis associated with platelet and monocyte adhesion and aggregation, atherosclerosis associated with smooth muscle proliferation and migration; restenosis, restenosis developed after thrombolysis therapies; restenosis developed after percutaneous transluminal angioplasties; restenosis developed after percutaneous transluminal coronary angioplasties and bypasses; inflammation; cardiovascular disease associated with metabolic syndrome, obesity, dyslipidemia, diabetes or high blood pressure; lipid-related disorders, dyslipidemia, hypercholesterolemia, hypertriglyceridemia, sitosterolemia, fatty liver disease, and hepatitis; preeclampsia; progression of polycystic kidney disease; and subcutaneous fat.

In other embodiments, the disease, condition or health disorder is selected from liver cirrhosis, liver cirrhosis associated with chronic liver disease, liver fibrosis, activation of hepatic stellate cells, accumulation of hepatic fibrous and total collagen; liver disease of necroinflammatory or immunological origin.

In other embodiments, the disease, condition or health condition is a disorder of the urogenital system selected from renal fibrosis; kidney failure resulting from chronic kidney disease or failure; kidney failure due to tissue accumulation or deposition and injury, progressive sclerosis, or glomerulonephritis; and prostate hypertrophy.

In further embodiments, the disease, condition or health condition is systemic sclerosis.

In other embodiments, the disease, condition or health condition is a cardiac disorder selected from cardiac interstitial fibrosis; cardiac remodeling and fibrosis and cardiac hypertrophy.

In other embodiments, the disease, condition or health condition is a CNS disorder or condition selected from Alzheimer's disease; Parkinson's disease; dementia; vascular cognitive impairment; cerebral vasospasms; and head trauma.

In other embodiments, the disease, condition or health condition is selected from ischemia or reperfusion injury; ischemia/reperfusion associated with organ transplant, lung transplant, lung transplant, or heart transplant; Conservation of blood substituents in trauma patients.

In further embodiments, the disease, condition or health disorder is a sexual, gynecological or urological disorder or condition selected from erectile dysfunction; impotence; premature ejaculation; female sexual dysfunction; female sexual desire dysfunction; hypoactive sexual desire disorder; vaginal atrophy, dyspaneuria, atrophic vaginitis; benign prostatic hyperplasia (BPH) or benign prostatic hypertrophy or enlargement; bladder outlet obstruction; painful bladder syndrome (BPS); interstitial cystitis (IC); overactive bladder syndrome, neurogenic bladder and incontinence; diabetic nephropathy.

In other embodiments, the disease, condition or health condition is selected from vaginal atrophy, dyspaneuria and atrophic vaginitis.

In other embodiments, the disease, condition or health condition is selected from benign prostatic hyperplasia (BPH) or benign prostatic hypertrophy or enlargement; bladder outlet obstruction; painful bladder syndrome (BPS); interstitial cystitis (IC); overactive bladder syndrome, neurogenic bladder and incontinence.

In further embodiments, the disease, condition or health disorder is a condition of a sexual nature, selected from erectile dysfunction; impotence; premature ejaculation; female sexual dysfunction; female sexual desire dysfunction and hypoactive sexual desire disorder.

In further embodiments, the disease or disorder is a diabetic nephropathy.

In further embodiments, the disease, condition or health disorder consists of Duchenne and Becker muscular dystrophies.

In other embodiments, the disease is an ocular disease or disorder selected from glaucoma, retinopathy, diabetic retinopathy, blepharitis, dry eye syndrome, and Sjogren's syndrome.

In further embodiments, the disease is a disease of the ear or a disorder selected from diminished hearing, partial loss or total loss of hearing; partial or total deafness; tinnitus; and noise-induced hearing loss.

In further embodiments, the disease is a topical or skin disorder or condition selected from dermal fibrosis, scleroderma and skin fibrosis.

In other embodiments, the treatment involves wound healing; wound healing in diabetics; improvement of microvascular perfusion; improved microvascular perfusion outcomes after injury; treatment of anal fissures; and treatment of diabetic ulcers.

In other embodiments, the disease or condition is selected from cancer metastases; osteoporosis; gastroparesis; functional dyspepsia; diabetic complications; diseases associated with endothelial dysfunction and neurological disorders associated with decreased nitric oxide production.

In another embodiment, the compounds of the invention can be provided in the form of implanted devices, such as stents. A stent is a mesh “tube” inserted into a natural passageway/duct in the body to prevent or counteract a disease-induced localized flow constriction. The term may also refer to a tube used to temporarily keep said natural canal open to allow access for surgery.

A drug-eluting stent (DES) is a peripheral or coronary stent (a base structure) placed in diseased and narrowed peripheral or coronary arteries, which slowly releases a drug to block cell proliferation, usually the proliferation of smooth muscle cells. This prevents fibrosis which, together with clots (thrombus), could block the artery carrying the stent, a process called restenosis. The stent is usually placed into the peripheral or coronary artery by a cardiology interventionist or radiology interventionist during an angioplasty procedure. Drugs commonly used in DES to block cell proliferation include analogs of paclitaxel or rapamycin.

In some embodiments of the invention, the compounds of claims 1 and 2 may be delivered via a drug eluting stent coated with said compounds. A drug eluting stent coated with the compounds of claims 1 and 2 may be useful for preventing restenosis and stent thrombosis during percutaneous coronary interventions. A drug eluting stent coated with the compounds of claims 1 and 2 may be capable of preventing the proliferation of smooth cells as well as facilitating revascularization and regeneration of the endothelial tissue of the artery into which the stent is inserted.

An alternative to percutaneous coronary intervention for the treatment of intractable angina due to occlusive coronary artery disease is a procedure called Coronary Artery Bypass Grafting (CABG). CABG provides only palliation of an ongoing process that is further complicated by the rapid development of graft atherosclerosis. The saphenous vein graft is the most commonly used conduit in CABG surgery. The long-term clinical success of venous CABG is hampered by three main reasons: accelerated graft atherosclerosis, incomplete endotalization, and thrombosis.

In some embodiments, the compounds of claims 1 and 2 can be used for the prevention of saphenous graft failure during CABG. The compound may aid in the endothelialization process and help prevent thrombosis. In this indication, the compounds of claims 1 and 2 are supplied locally in the form of a gel.

The terms, “disease,” “disorder,” and “condition” may be used interchangeably herein in reference to a medical or pathological condition mediated by sGC, cGMP, and/or NO.

As used herein, the terms “individual” and “patient” are used interchangeably. The terms “individual” and “patient” refer to an animal (for example, a bird such as a chicken, quail or turkey, or a mammal), specifically a “mammal”, including a non-primate (for example, a cow). , pig, horse, sheep, rabbit, guinea pig, rat, cat, dog, and mouse) and a primate (for example, a monkey, chimpanzee, and a human) and, more specifically, a human. In some embodiments, the individual is a non-human animal, such as a farm animal (e.g., a horse, cow, pig, or sheep) or a pet (e.g., a dog, cat, guinea pig, or rabbit). In some embodiments, the subject is a human being.

As used herein, the term “biological sample” refers to an in vitro or ex vivo sample, and includes, without limitation, cell cultures or extracts thereof; biopsied material obtained from or extracts from a mammal; blood, saliva, urine, feces, semen, tears, lymphatic fluid, ocular fluid, vitreous humor, or other body fluids or extracts thereof.

“Treat”, “treating” or “treatment” with respect to a disorder or disease means the alleviation or elimination of the cause and/or effects of the disorder or disease. As used herein, the terms "treat", "treatment" and "treating" refer to the reduction or improvement of the progression, severity and/or duration of a condition mediated by sGC, cGMP and/or NO, or the improvement of one or more symptoms (preferably, one or more discernible symptoms) of said condition (i.e., "control" without "curing" the condition), as a result of the administration of one or more therapies (for example, one or plus therapeutic agents, such as a compound or composition of the invention). In specific embodiments, the terms “treat”; “Treatment” and “treating” refer to the improvement of at least one measurable physical parameter of a condition mediated by sGC, cGMP and/or NO. In other embodiments, the terms "treat", "treatment" and "treating" refer to inhibiting the progression of a condition mediated by sGC, cGMP and/or NO, either physically by, for example, stabilizing a symptom. discernible or physiologically through, for example, the stabilization of a physical parameter, or both.

As used herein, the term "avoid" refers to pre-administering a drug to avoid or prevent the appearance of one or more symptoms of a disease or disorder. The expert in medical technique understands that the term “prevent” is not an absolute term. In the medical industry it is understood to refer to the prophylactic administration of a drug to substantially reduce the likelihood or severity of a condition or a symptom of the condition and this is the intended meaning of this description. In The Physician's Desk Reference, a standard text in the field, the term “prevent” is used hundreds of times. As used herein, the terms "prevent", "preventing" and "prevention" with respect to a disorder or disease, refer to avoiding the cause, effects, symptoms or progression of a disease or disorder before the disease or disorder itself manifests itself completely.

The compounds and pharmaceutical compositions described herein can be used alone or in combination therapy for the treatment or prevention of a disease or disorder mediated, regulated or influenced by sGC, cgMp and/or NO.

The compounds and compositions described herein are also useful for the veterinary treatment of companion animals, exotic animals and farm animals, including, without limitation, dogs, cats, mice, rats, hamsters, gerbils, guinea pigs, rabbits, horses. , pigs and cattle.

Combination therapies

The compounds and pharmaceutical compositions described herein can be used in combination therapies with one or more additional therapeutic agents. For combination treatment with more than one active agent, where the active agents are in separate dosage formulations, the active agents may be administered separately or together. Furthermore, the administration of one element may take place before, at the same time as, or after the administration of the other agent.

When administered together with other agents, for example, when administered together with another pain drug, an “effective amount” of the second agent will depend on the type of drug used. Suitable doses are known for the approved agents and can be adjusted by one skilled in the art, depending on the condition of the individual, the type of condition or conditions being treated and the amount of a compound described in the present description that is being used. In cases where no quantity is expressly indicated, an effective quantity must be assumed. For example, the compounds described herein can be administered to an individual with a dosage range of about 0.01 to about 10,000 mg/kg body weight/day, from about 0.01 to about 5,000 mg/kg body weight body weight/day, from about 0.01 to about 3,000 mg/kg body weight/day, from about 0.01 to about 1,000 mg/kg body weight/day, from about 0.01 to about 500 mg/kg body weight/day, from about 0.01 to about 300 mg/kg body weight/day, from about 0.01 to about 100 mg/kg body weight/day.

When "combination therapy" is employed, an effective amount can be achieved using a first amount of a compound of claims 1 and 2 or a pharmaceutically acceptable salt thereof and a second amount of a suitable additional therapeutic agent.

In one embodiment of this invention, a compound of claims 1 and 2 and the additional therapeutic agent are each administered in an effective amount (i.e., each in an amount that would be therapeutically effective if administered alone). In another embodiment, a compound of claims 1 and 2 and the additional therapeutic agent are each administered in an amount that does not itself provide a therapeutic effect (a subtherapeutic dose). In another embodiment, a compound of claims 1 and 2 may be administered in an effective amount, while the additional therapeutic agent is administered in a subtherapeutic dose. In another embodiment, a compound of claims 1 and 2 may be administered in a subtherapeutic dose, while the additional therapeutic agent, for example, a suitable cancer therapeutic agent, is administered in an effective amount.

As used herein, the terms “in combination” or “coadministration” may be used interchangeably in reference to the use of more than one therapy (e.g., one or more prophylactic and/or therapeutic agents). Use of the terms does not limit the order in which therapies (e.g., prophylactic and/or therapeutic agents) are administered to an individual.

Coadministration encompasses the administration of the first and second amounts of the compounds in a substantially simultaneous manner, such as in a single pharmaceutical composition, for example, a capsule or tablet having a fixed ratio of first and second amounts, or in multiple separate capsules or tablets for each one. Furthermore, such coadministration also encompasses the use of each compound sequentially in any order. When coadministration involves the separate administration of the first amount of a compound of claims 1 and 2 and a second amount of an additional therapeutic agent, the compounds are administered sufficiently close together in time to obtain the desired therapeutic effect. For example, the period of time between each administration that can result in the desired therapeutic effect can vary from minutes to hours and can be determined taking into account the properties of each compound, such as potency, solubility, bioavailability, plasma half-life and profile. kinetic. For example, a compound of claims 1 and 2 and the second therapeutic agent can be administered in any order at an interval of about 24 hours apart, at an interval of about 16 hours apart, at an interval of about 8 hours apart, an interval of approximately 4 hours from each other, an interval of approximately 1 hour from each other or at an interval of approximately 30 minutes from each other.

More specifically, a first therapy (e.g., a prophylactic or therapeutic agent such as a compound described herein) may be administered before (e.g., 5 minutes, 15 minutes, 30 minutes, 45 minutes, 1 hour, 2 hours, 4 hours, 6 hours, 12 hours, 24 hours, 48 hours, 72 hours, 96 hours, 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 8 weeks, or 12 weeks before), simultaneously with, or after (e.g., 5 minutes, 15 minutes, 30 minutes, 45 minutes, 1 hour, 2 hours, 4 hours, 6 hours, 12 hours, 24 hours, 48 hours, 72 hours, 96 hours, 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 8 weeks, or 12 weeks later) the administration of a second therapy (for example, a prophylactic or therapeutic agent such as an anticancer agent) to a individual.

Examples of other therapeutic agents that may be combined with a compound of this description, whether administered separately or in the same pharmaceutical composition include, but are not limited to:

(1) Endothelium-derived releasing factor (EDRF);

(2) NO donors such as a nitrosothiol, a nitrite, a sidnoniimine, a NONOate, an N-nitrosamine, an N-hydroxylnitrosamine, a nitrosamine, nitrotyrosine, a diazetine dioxide, an oxatriazole 5-amine, an oxime, a hydroxylamine, an N-hydroxyguanidine, a hydroxyurea or a furoxane. Some examples of these types of compounds include: glyceryl trinitrate (also known as GTN, glyceryl nitrate, and trinitrogylcerin), the nitrate ester of glycerol; sodium nitroprusside (SNP), where a nitric oxide molecule coordinates with iron metal forming a square dipyramidal complex; 3-morpholinosidnonimine (SIN-1), a zwitterionic compound formed by the combination of a morpholine and a sydnonimine; S-nitroso-N-acetylpenicillamine (SNAP), an N-acetylated amino acid derivative with a nitrosothiol functional group; diethylenetriamine/NO (DETA/NO), a nitric oxide compound covalently linked to diethylenetriamine; and NCX 4016, an m-nitroxymethyl phenyl ester of acetylsalicylic acid. More specific examples of some of these classes of NO donors include: classical nitrovasodilators, such as organic nitrate and nitrite esters, including nitroglycerin, amyl nitrite, isosorbide dinitrate, isosorbide 5-mononitrate and nicorandyl; Isosorbide (Dilatrate®-SR, Imdur®, Ismo®, Isordil®, Isordil®, Titradose®, Monoket®), FK 409 (NOR-3); FR 144420 (NOR-4); 3-morpholinosidnonimine; linsidomine hydrochloride (“SIN-1”); S-nitroso-N-acetylpenicillamine (“SNAP”); AZD3582 (leading compound CINOD), NCX 4016, NCX 701, NCX 1022, HCT 1026, NCX 1015, NCX 950, NCX 1000, NCX 1020, AZD 4717, NCX 1510/NCX 1512, NCX 2216, and NCX 4040 (all marketed by NicOx S.A.), S-nitrosoglutathione (GSNO), sodium nitroprusside, S-nitrosoglutathione mono-ethyl ester (GSNO-ester),6-(2-hydroxy-1-methyl-nitrosohydrazine)-W-methyl-1- hexanamine (NOC-9) or diethylamine NONOate. Nitric oxide donors are also described in US-5,155,137, US-5,366,997, US-5,405,919, US-5,650,442, US-5,700,830, US-5,632,981, US-6,290. 981, US-5,691,423, US-5,721,365, US-5,714,511, US-6,511,911 and US-5,814,666, Chrysselis et al. (2002) J Med Chem. 45:5406-9 (such as NO donors 14 and 17), and Nitric Oxide Donors for Pharmaceutical and Biological Research, eds: Peng George Wang, Tingwei Bill Cai, Naoyuki Taniguchi, Wiley, 2005 ;

(3) Other substances that improve cGMP concentrations, such as protoporphyrin IX, arachidonic acid, and phenylhydrazine derivatives;

(4) nitric oxide synthase substrates: for example, n-hydroxyguanidine-based analogues, such as N[G]-hydroxy-L-arginine (NOHA), 1-(3,4-dimethoxy-2-dorobenzylidenamino)-3 -hydroxyguanidine and PR5 (1-(3,4-dimethoxy-2-dorobenzylidenamino)-3-hydroxyguanidine); L-arginine derivatives (such as homo-Arg, homo-NOHA, N-tert-butyloxy- and N-(3-methyl-2-butenyl)oxy-L-arginine, canavanine, epsilon guanidine-carpoic acid, agmatine, hydroxyl-agmatine and L-tyrosyl-L-arginine)); N-alkyl-N'-hydroxyguanidines (such as N-cyclopropyl-N'-hydroxyguanidine and N-butyl-N'-hydroxyguanidine), N-aryl-N'-hydroxyguanidines (such as N-phenyl-N'-hydroxyguanidine and their para-substituted derivatives bearing the substituents -F, -Cl, -methyl, -OH, respectively); guanidine derivatives, such as 3-(trifluormethyl)propylguanidine; and others indicated by Cali et al. (2005, Current Topics in Medicinal Chemistry 5:721-736) and described in the references cited therein;

(5) Compounds that enhance eNOS transcription: for example, those described in WO 02/064146, WO 02/064545, WO 02/064546 and WO 02/064565 and corresponding patent documents, such as US2003/0008915, US2003/0022935 , US2003/0022939 and US2003/0055093. Other eNOS transcriptional enhancers, including those described in US20050101599 (e.g., 2,2-difluorobenzo[1,3]dioxol-5-carboxylic acid indan-2-ylamide, and 4-fluoro-N-(indan-2-yl )-benzamide), and the Sanofi-Aventis compounds AVE3085 and AVE9488 (CA Registry NO. 916514 70-0; Schafer et al., Journal of Thrombosis and Homeostasis 2005; volume 3, supplement 1: abstract number P1487);

(6) NO-independent heme-independent sGC activators including, but not limited to: BAY 58-2667 (see patent publication DE19943635)

HMR-1766 (ataciguat sodium, see patent publication WO2000002851)

S 3448 (2-(4-chloro-phenylsulfonylamino)-4,5-dimethoxy-N-(4-(thiomorpholine-4-sulfonyl)-phenyl)-benzamide (see patent publications DE19830430 and WO2000002851)

and HMR-1069 (Sanofi-Aventis).

(7) Heme-dependent GC stimulators including, but not limited to:

YC-1 (see patent publications EP667345 and DE19744026)

Riociguat (BAY 63-2521, Adempas, commercial product, described in DE19834044)

Neliciguat (BAY 60-4552, described in WO 2003095451)

Vericiguat (BAY 1021189, clinical alternative to Riociguat),

BAY 41-2272 (described in DE19834047 and DE19942809)

BAY 41-8543 (described in DE19834044)

Etriciguat (described in WO 2003086407)

CFM-1571 (see patent publication WO2000027394)

A 344905, its acrylamide analogue A-350619 and the aminopyrimidine analogue A-778935.

A-344905;

A-778935;

Compounds described in one of the publications: US20090209556, US8455638, US20110118282 (WO2009032249), US20100292192, US20110201621, US7947664, US8053455 (WO2009094242), 16764, US8507512, (WO2010099054) US20110218202 (WO2010065275), US20130012511 (WO2011119518), US20130072492 (WO2011149921), US20130210798 (WO2012058132) and other compounds described in Tetrahedron Letters (2003), 44(48): 8661-8663.

(8) Compounds that inhibit cGMP degradation, such as:

PDE5 inhibitors, such as, for example, Sildenafil (Viagra®) and other related agents, such as Avanafil, Lodenafil, Mirodenafil, sildenafil citrate (Revatio®), Tadalafil (Cialis® or Adcirca®), Vardenafil (Levitra®) and Udenafil; Alprostadil; and Dipyridamole;

(9) Calcium channel blockers, such as:

Dihydropyridine calcium channel blockers: Amlodipine (Norvasc), Aranidipine (Sapresta), Azelnidipine (Calblock), Barnidipine (HypoCa), Benidipine (Coniel), Cilnidipine (Atelec, Cinalong, Siscard), Clevidipine (Cleviprex), Diltiazem, Efonidipine (Landel), Felodipine (Plendil), Lacidipine (Motens, Lacipil), Lercanidipine (Zanidip), Manidipine (Calslot, Madipine), Nicardipine (Cardene, Carden SR), Nifedipine (Procardia, Adalat), Nilvadipine (Nivadil), Nimodipine ( Nimotop), Nisoldipine (Baymycard, Sular, Syscor), Nitrendipine (Cardif, Nitrepin, Bailotensin), Pranidipine (Acalas), Isradipine (Lomir);

Phenylalkylamine calcium channel blockers: Verapamil (Calan, Isoptin)

Galopamyl (Procorum, D600);

Benzothiazepines: Diltiazem (Cardizem);

Non-selective calcium channel inhibitors such as: mibefradil, bepridil and fluspirilene, fendiline;

(10) Endothelin receptor antagonists (ERAs): for example, the dual endothelin receptor antagonist (ET<a>and ET<b>) Bosentan (marketed as Tracleer®); Sitexentan, marketed under the name Thelin®; Ambrisentan is marketed as Letairis® in the US; dual/nonselective endothelin antagonist Actelion-1, which entered clinical trials in 2008;

(11) Prostacyclin derivatives or analogues: for example, prostacyclin (prostaglandin I2), Epoprostenol (synthetic prostacyclin, marketed as Flolan®); Treprostinil (Remodulin®), Iloprost (Ilomedin®), Iloprost (marketed as Ventavis®); oral and inhaled forms of Remodulin® that are under development; Beraprost, an oral prostanoid available in Japan and South Korea;

(12) Antihyperlipidemics, such as: bile acid sequestrants (for example, Cholestyramine, Colestipol, Colestilane and Colesevelam); statins, such as Atorvastatin, Simvastatin, Lovastatin, Fluvastatin, Pitavastatin, Rosuvastatin and Pravastatin; cholesterol absorption inhibitors such as Ezetimibe; other lipid-lowering agents, such as Icosapent ethyl ester, omega-3 fatty acid ethyl esters, Reducol; fibric acid derivatives such as Clofibrate, Bezafibrate, Clinofibrate, Gemfibrozil, Ronifibrate, Binifibrate, Fenofibrate, Ciprofibrate, Choline fenofibrate; nicotinic acid derivatives such as Acipimox and Niacin; also combinations of statins, niacin, intestinal cholesterol absorption inhibition supplements (ezetimib and others) and fibrates; antiplatelet therapies, such as clopidogrel bisulfate;

(13) Anticoagulants, such as the following types:

• Coumarins (Vitamin K antagonists): Warfarin® (Coumadin) used mainly in the United States and the United Kingdom; Acenocoumarol® and Phenprocoumon®, mainly used in other countries Fenindione®; • Heparin and derived substances, such as: Heparin; low molecular weight heparin, Fondaparinux and Idraparinux;

• direct thrombin inhibitors such as: Argatroban, Lepirudin, Bivalirudin and Dabigatran; Ximelogatran (Exanta®) not approved in the United States;

• Tissue plasminogen activators, used to dissolve clots and unblock arteries, such as alteplase; (14) Antiplatelet drugs: for example, thienopyridines, such as Lopidogrel and Ticlopidine; Dipyridamole; Aspirin; (15) ACE inhibitors, for example, the following types:

• Sulfhydryl-containing agents, such as Captopril (trade name Capoten®), the first ACE inhibitor, and Zofenpril;

• Dicarboxylate-containing agents, such as Enalapril (Vasotec/Renitec®); Ramipril (Altace/Tritace/Ramace/Ramiwin®); Quinapril (Accupril®), Perindopril (Coversil/Aceon®); Lisinopril (Lisodur/Lopril/Novatec/Prinivil/Zestril®) and Benazepril (Lotensin®);

• Phosphonate-containing agents such as: Fosinopril;

• Naturally occurring ACE inhibitors, such as: Casokinins and lactokinins, which are breakdown products of casein and whey that occur naturally after ingestion of dairy products, especially cultured milk; The lactotripeptides Val-Pro-Pro and Ile-Pro-Pro produced by the casein-derived probiotic Lactobacillus helveticus also have ACE inhibitory functions and antihypertensive functions;

• Other ACE inhibitors, such as Alacepril, Delapril, Cilazapril, Imidapril, Trandolapril, Temocapril, Moexipril, Spirapril,

(16) Oxygen supplement therapy;

(17) Beta blockers, such as the following types:

• Non-selective agents: Alprenolol®, Bucindolol®, Carteolol®, Carvedilol® (has additional a-blocking activity), Labetalol® (has additional a-blocking activity), Nadolol®, Penbutolol® (has intrinsic sympathomimetic activity), Pindolol® (has intrinsic sympathomimetic activity), Oxprenonol, Acebutolol, Sotalol, Mepindolol, Celiprolol, Arotinolol, Tertatolol, Amosulalol, Nipradilol, Propranolol® and Timolol®;

• p-i-Selective agents: Acebutolol® (has intrinsic sympathomimetic activity), Atenolol®, Betaxolol®, Bisoprolol®, Celiprolol®, dobutamine hydrochloride, irsogladine maleate, Carvedilol, Talinolol, Esmolol®, Metoprolol® and Nebivolol®;

• p2-Selective agents: Butaxamine® (slight a-adrenergic agonist activity);

(18) Antiarrhythmic agents such as the following types:

• Type I (sodium channel blockers): Quinidine, Lidocaine, Phenytoin, Propafenone

• Type III (potassium channel blockers): Amiodarone, Dofetilide, Sotalol

• Type V: Adenosine, Digoxin

(19) Diuretics such as: Thiazide diuretics, for example, Chlorothiazide, Clothalidone and Hydrochlorothiazide, Bendroflumethiazide, Cyclopenthiazide, Methychlothiazide, Polythiazide, Quinetaazone, Xipamide, Metolazone, Indapamide, Cycletanine; Loop diuretics, such as Furosemide and Toresamide; potassium-sparing diuretics, such as Amiloride, Spironolactone, Potassium Canrenoate, Eplerenone and Triamterene; combinations of these agents; other diuretics such as Acetazolamide and Carperitide

(20a) Direct acting vasodilators, such as hydralazine hydrochloride, diazoxide, sodium nitroprusside, cadralazine; other vasodilators such as isosorbide and isosorbide 5-mononitrate;

(20b) Exogenous vasodilators, such as

• Adenocard®, an adenosine agonist, primarily used as an antiarrhythmic;

• Alpha blockers (which block the vasoconstrictive effect of adrenaline):

Alpha-1 adrenoceptor antagonists such as Prazosin, Indoramin, Urapidil, Bunazosin, Terazosin, Doxazosin

• Atrial natriuretic peptide (ANP);

• Ethanol;

• Histamine inducers, which complement C3a, C4a, and C5a proteins, act by activating histamine release from mast cells and basophilic granulocytes;

• Tetrahydrocannabinol (THC), the main active chemical substance in marijuana that has slightly vasodilatory effects;

• Papaverine, an alkaloid found in papaver somniferum, the opium poppy;b

(21) Bronchodilators: There are two main types of bronchodilators, p2 agonists and anticholinergics, illustrated below:

• p2 agonists: Salbutamol® or albuterol (common trade name: Ventolin) and Terbutaline® are fast-acting p2 agonists for rapid relief of COPD symptoms. Long-acting p2 agonists (LABA) such as Salmeterol® and Formoterol®;

• anticholinergics: Ipratropium® is the most widely prescribed fast-acting anticholinergic drug. Tiotropium® is the most widely prescribed long-acting anticholinergic drug for COPD;

• Theophylline®, a bronchodilator and phosphodiesterase inhibitor;

(22) Corticosteroids: such as beclomethasone, methylprednisolone, betamethasone, prednisone, prednisolone, triamcinolone, fexamethasone, fluticasone, flunisolide and hydrocortisone, and corticosteroid analogues such as budesonide

(23) Dietary supplements such as, for example: omega-3 oils; folic acid, niacin, zinc, copper, Korean red ginseng, ginkgo, pine bark, Tribulus terrestris, arginine, Avena sativa, Epimedium, maca root, muira puama, Serenoa repens, and Swedish flower pollen; vitamin C, vitamin E, vitamin K2; Testosterone supplements, testosterone transdermal patch; Zoraxel, Naltrexone, Bremelanotide (formerly PT-141), Melanotan II, hMaxi-K; Prelox: a proprietary blend/combination of natural ingredients, L-arginine aspartate and pycnogenol;

(24) PGD2 receptor antagonists including, but not limited to, compounds of PGD2 antagonist activity described in United States Patent Applications US20020022218, US20010051624, and US20030055077, PCT Published Applications W09700853, W09825919, WO03066046, 66047, WO03101961 WO03101981, WO04007451, WO0178697, WO04032848, WO03097042, WO03097598, WO03022814, WO03022813, and WO04058164, European patent applications indicated in: Torisuycol. 2004Bioorg Med Chem Lett14:4557, Torisuy al. 2004 BioorgMed Chem Lett200414:4891, and Torisu et al. 2004Bioorg & Med Chem2004 12:4685;

(25) Immunosuppressants such as cyclosporine (cyclosporin A, Sandimmune® nebucal®), tacrolimus (FK-506, Prograf®), rapamycin (sirolimus, Rapamune®) and other FK-506 type immunosuppressants and mycophenolate, e.g. mofetil mycophenolate (CellCept®);

(26) Nonsteroidal asthmatic antiasthmatics such as p2 agonists (e.g., terbutaline, metaproterenol, fenoterol, isoetarine, albuterol, salmeterol, bitolterol and pirbuterol) and p2 agonist-corticosteroid combinations (e.g., salmeterol-fluticasone (Advair®), formoterol-budesonide (Symbicort®)), theophylline,cromoline,cromoline sodium, nedocromil, atropine, ipratropium, ipratropium bromide, leukotriene biosynthesis inhibitors (zileuton, BAY1005);

(27) Nonsteroidal anti-inflammatory agents (NSAIDs), such as propionic acid derivatives (e.g., alminoprofen, benoxaprofen, bucloxic acid, carprofen, fenbufen, fenoprofen, fluprofen, flurbiprofen, ibuprofen, indoprofen, ketoprofen, miroprofen, naproxen, oxaprozin, pirprofen, pranoprofen, suprofen, tiaprofenic acid and thioxaprofen), acetic acid derivatives (e.g. indomethacin, acematacin, alclofenac, clidanac, diclofenac, fenclofenac, phenclozic acid, fentiazac, furofenac, ibufenac, isoxepaco, oxpinac, sulindac, thiopinac, tolmetin , zidomethacin and zomepirac), fenamic acid derivatives (for example, flufenamic acid, meclofenamic acid, mefenamic acid, niflumic acid and tolfenamic acid), biphenylcarboxylic acid derivatives (for example, diflunisal and flufenisal), oxicams (for example, isoxicam, piroxicam, sudoxicam and tenoxicam), salicylates (for example, acetylsalicylic acid and sulfasalazine) and pyrazolones (for example, apazone, bezpiperilone, feprazone, mofebutazone, oxyphenbutazone and phenylbutazone);

(28) Cyclooxygenase-2 (COX-2) inhibitors, such as celecoxib®(Celebrex), rofecoxib®(Vioxx), valdecoxib, etoricoxib, parecoxib and lumiracoxib;

(opioid analgesics such as codeine, fentanyl, hydromorphone, levorphanol, meperidine, methadone, morphine, oxycodone, oxymorphone, propoxyphene, buprenorphine, butorphanol, dezocine, nalbuphine and pentazocine; and

(29) Antidiabetic agents such as insulin and insulin mimetics, sulfonylureas (e.g. Glyburide, Glibenclamide, Glipizide, Gliclazide, Gliquidone, Glimepiride, Meglinatide, Tolbutamide, Chlorpropamide, Acetohexamide, Tolazamide), biguanides, e.g. metformin (Glucophage® ), α-glucosidase inhibitors (such as Acarbose, Epalrestat, Voglibosa, Miglitol), thiazolidinone compounds, for example, rosiglitazone (Arandia®), troglitazone (Rezulin®), ciglitazone, pioglitazone (Actos®) and englitazone; insulin sensitizers such as Pioglitazone and Rosiglitazone; insulin secretagogues such as Repaglinide, Nateglinide and Mytilinide; incretin mimetics such as Exanatide and Liraglutide; amylin analogues such as Pramlintide; glucose lowering agents such as chromium picolinate (optionally combined with biotin); dipeptidyl peptidase IV inhibitors such as sitagliptin, vildagliptin, saxagliptin, alogliptin and linagliptin; vaccines currently in development for the treatment of diabetes; AVE-0277, Alum-GAD, Bh T-3021, IBC-VS01; cytokine-targeted therapies in development for the treatment of diabetes such as Anakinra, Canakinumab, Diacerein, Gevokizumab, LY-2189102, MABP-1, GIT-027; drugs in development for the treatment of diabetes:

(30) HDL cholesterol-increasing agents, such as Anacetrapib, MK-524, CER-001, DRL-17822, Dalcetrapib, JTT-302, RVX-000222, TA-8995;

(31) Anti-obesity drugs such as methamphetamine hydrochloride, amfepramone hydrochloride (Tenuate®), Phentermine (Ionamin®), benzphetamine hydrochloride (Didrex®), phendimetrazine tartrate (Bontril®, Prelu-2®, Plegine®) , Mazindol (Sanorex®), Orlistat (Xenical®), Sibutramine hydrochloride monohydrate (Meridia®, Reductil®), Rimonabant (Acomplia®), Amfepramone, chromium picolinate, RM-493, TZP-301; combination such as phentermine/topiramate, bupropion/naltrexone, sibutramine/metformin, bupropion SR/zonisamide SR, salmeterol, xinafoate/fluticasone propionate; lorcaserin hydrochloride hydrochloride, phentermine/topiramate, bupropion/naltrexone, cetilistat, exenatide, KI-0803, liraglutide, metformin hydrochloride, sibutramine/metformin, 876167, ALS-L-1023, bupropion SR/zonisamide SR, CORT-108297, canagliflozin , chromium picolinate, GSK-1521498, LY-377604, metreleptin, obinepitide, P-57AS3, PSN-821, salmeterol xinafoate/fluticasone propionate, sodium tungstate, somatropin (recombinant), TM-30339, TTP-435, tesamorelin, tesofensine, velneperite, zonisamide, BMS-830216, ALB-127158, AP-1030, ATHX-105, AZD-2820, AZD-8329, beloranib hemioxalate, CP-404, HPP-404, ISIS-FGFR4Rx, insulinotropin, KD-3010PF, 05212389, PP-1420, PSN-842, Peptide YY3-36, Resveratrol, S-234462; S-234462, sobetiroma, TM-38837, tetrahydrocannabivarin, ZYO-1, beta-lapachone;

(32) angiotensin receptor blockers such as losartan, valsartan, candesartan cilexetil, eprosartan, irbesartan, telmisartan, olmesartan medoxomil, azilsartan medoxomil;

(33) Renin inhibitors such as Aliskiren hemifumarate;

(34) Centrally acting alpha-2-adrenoceptor agonists, such as methyldopa, clonidine, guanfacine;

(35) Adrenergic neuronal blockers, such as guanethidine, guanadrel;

(36) Imidazoline I-1 receptor agonists, such as rimenidine hydrogen phosphate and moxonidine hydrochloride hydrate;

(37) Aldosterone antagonists such as spironolactone and eplerenone

(38) Potassium channel activators such as pinacidil

(39) Dopamine D1 agonists, such as fenoldopam mesylate; Other dopamine agonists, such as ibopamine, dopexamine and docarpamine;

(40) 5-HT2 antagonists, such as katanserin;

(41) Drugs currently in development for the treatment of high blood pressure:

(42) Vasopressin antagonists such as Tolvaptan;

(43) Calcium channel sensitizers such as Levosimendan or activators such as Nicorandil;

(44) PDE-3 inhibitors such as Amrinone, Milrinone, Enoximone, Vesnarinone, Pimobendan, Olprinone; (45) Adenylate cyclase activators, such as Colforsin dapropate hydrochloride;

(46) Positive inotropic agents such as Digoxin and Methyldigoxin; cardiotonic metabolic agents, such as Ubidecarenone; brain natriuretic peptides, such as Nesiritide;

(47) Drugs currently in development for the treatment of heart failure:

(48) Drugs currently in development for the treatment of pulmonary hypertension:

(49) Drugs currently in development for the treatment of female sexual dysfunction:

(50) Drugs used for the treatment of erectile dysfunction, such as Alprostadil, Aviptadil, phenylamine mesylate, Weige, Alprostadil;

(51) Drugs currently in development for the treatment of male sexual dysfunction:

(51) Drugs in development for the treatment of sleep apnea:

(52) Drugs currently in development for the treatment of metabolic syndrome:

(53) Anti-obesity drugs:

(54) Medications used to treat Alzheimer's disease: for example, cholinesterase inhibitors prescribed for mild to moderate Alzheimer's disease, including Razadyne® (galantamine), Exelon® (rivastigmine), and Aricept® (donepezil), Cognex® (tacrine); Namenda® (memantine), an antagonist of N-methyl D-aspartate (NMDA) and Aricept®, prescribed to treat moderate to severe Alzheimer's disease; vitamin E (an antioxidant).

(55) Antidepressants: tricyclic antidepressants such as amitriptyline (Elavil®), desipramine (Norpramin®), imipramine (Tofranil®), amoxapine (Asendin®), nortriptyline; selective serotonin reuptake inhibitors (SSRIs) such as paroxetine (Paxil®), fluoxetine (Prozac®), sertraline (Zoloft®), and citralopram (celexa®); and others such as doxepin (sinecuan®) and trazodone (desarrel®); SNRI (for example, venlafaxine and reboxetine); dopaminergic antidepressants (for example, bupropione and amineptine).

(56) Neuroprotective agents: for example, memantine, L-dopa, bromocriptine, pergolide, talipexole, pramipexole, cabergoline, neuroprotective agents currently under investigation, including antiapoptotic drugs (CEP 1347 and CTCT346), lazaroids, bioenergetics, antiglutamatergic agents and receptors dopamine. Other clinically evaluated neuroprotective agents are, for example, the monoamine oxidase B inhibitors selegiline and rasagiline, dopamine agonists, and the mitochondrial complex I enhancer coenzyme Q10.

(57) Antipsychotic medications: for example, ziprasidone (Geodon™), risperidone (Risperdal™), and olanzapine (Zyprexa™).

Kits

The pharmaceutical compounds and formulations described herein may be contained in a kit. The kit may include single or multiple doses of two or more agents, each packaged or formulated individually, or single or multiple doses of two or more agents packaged or formulated in combination. Therefore, one or more agents may be present in the first container, and the kit may optionally include one or more agents in a second container. The container or containers are placed within a container, and the container may optionally include administration or dosage instructions. A kit may include additional components, such as syringes or other means for administering the agents, as well as diluents or other means for formulation. Therefore, the kits may comprise: a) a pharmaceutical composition comprising a compound described in the present description and a pharmaceutically acceptable carrier, vehicle or diluent; and b) a container or container. The kits may optionally comprise instructions describing a method of using the pharmaceutical compositions in one or more of the methods described herein (for example, preventing or treating one or more of the diseases and disorders described herein). . The kit may optionally comprise a second pharmaceutical composition comprising one or more additional agents described herein for use in adjuvant therapy, a pharmaceutically acceptable carrier, vehicle or diluent. The pharmaceutical composition comprising the compound described in the present description and the second pharmaceutical composition contained in the kit may optionally be combined into the same pharmaceutical composition.

A kit includes a container or container for containing the pharmaceutical compositions and may also include divided containers such as a divided bottle or a divided foil package. The container may be, for example, a paper or cardboard box, a glass or plastic bottle or vial, a resealable bag (for example, to contain a “spare” tablet to be placed in a different container), or a container. bubble or blister with individual doses to press out of the package according to a therapeutic program. It is feasible that more than one container may be used together in a single package to market a single dosage form. For example, the tablets may be contained in a bottle which is in turn contained within a box.

An example of a kit is the so-called bubble or blister packaging. Bubble or blister packaging is well known in the packaging industry and is widely used for packaging unit dosage forms (tablets, capsules, and the like). Bubble or blister packaging generally consists of a sheet of relatively rigid material covered with a sheet of a preferably transparent plastic material. During the packaging process, cavities form in the plastic sheet. The cavities are sized and shaped as individual tablets or capsules for packaging or may be sized and shaped to accommodate multiple tablets and/or capsules for packaging. The tablets or capsules are then placed in the corresponding cavities and the sheet of relatively rigid material is sealed against the plastic sheet on the side of the sheet opposite the direction in which the cavities were formed. Accordingly, the tablets or capsules are sealed individually or together, as desired, in the cavities between the plastic sheet and the sheet. Preferably the strength of the film is such that the tablets or capsules can be removed from the bubble or blister pack by manually applying pressure to the cavities so that an opening is formed in the film at the location of the cavity. The tablet or capsule can then be removed through the opening.

It may be desirable to provide a written means of avoiding forgetfulness that contains information and/or instructions to the doctor, pharmacist or individual about when to take the medication. A “daily dose” may be a single tablet or capsule or several tablets or capsules to take on a given day. When the kit contains separate compositions, a daily dose of one or more kit compositions may consist of one tablet or capsule, while a daily dose of another or more kit compositions may consist of several tablets or capsules. A kit may take the form of a dispenser designed to dispense daily doses at a time in their intended order of use. The dispenser can be equipped with a means to prevent forgetfulness to further facilitate compliance with the regimen. An example of such a means of avoiding forgetfulness is a mechanical counter that indicates the number of daily doses that have been dispensed. Another example of such a means of preventing forgetfulness is a battery-powered microchip memory coupled with a liquid crystal readout or audible reminder signal that, for example, reads the date the last daily dose has been taken and/or remembers when the last daily dose was taken. you should take the next dose.

Examples

As used in the present description, all abbreviations, symbols and conventions are in accordance with those used in contemporary scientific literature. See, for example, Janet S. Dodd, ed., The ACS Stile Guide: A Manual for Authors and Editors, 2nd ed., Washington, D.C.: American Chemical Society, 1997.

Example 1: Synthesis of the compounds of Table 1A, Table 1B, Table 1C and Table 1D.

General procedure A

Dione enolate formation: To a solution of ketone A in THF cooled to -78 °C, LiHMDS (e.g., 0.9 equiv, 1.0 M in toluene) was added dropwise via syringe. The reaction was allowed to warm to 0 °C, then charged with diethyl oxalate (1.2 equiv). At that time, the reaction was warmed to room temperature and stirred at this temperature until it was considered complete (e.g., using TLC or LC/MS analysis). Once the reaction was complete (reaction time was typically 45 minutes), the dione enolate B product was used “as is” in step 2, i.e., the cyclization step, without further purification.

Stage 2:

Pyrazole formation: Dione enolate B was diluted with ethanol and consecutively loaded with HCl (e.g., 3 equiv., 1.25 M solution in ethanol) and arylhydrazine hydrate (e.g., 1.15 equiv.). The reaction mixture was heated to 70°C and stirred at this temperature until cyclization was considered complete (e.g., by LC/Ms analysis, typically 30 minutes). Once complete, the reaction mixture was carefully treated with solid sodium bicarbonate (e.g., 4 equiv.) and diluted with dichloromethane and water. The layers were separated and the aqueous layer was further diluted with water before extraction with dichloromethane (3 times). The combined organic layers were washed with brine, dried over MgSO4, filtered, and concentrated in vacuo. The resulting pyrazole C was then purified by SiO2 chromatography using an appropriate gradient of EtOAc in hexanes.

Stage 3:

Formation of amidine: To a suspension of NH4Cl (e.g., 5 equiv.) in toluene cooled to 0 °C, AlMe3 (e.g., 5 equiv., 2.0 M solution in toluene) was added dropwise with a syringe. . The reaction was allowed to warm to room temperature and stirred at this temperature until no further bubbling was observed. Pyrazole C was added in 1 part to the reaction mixture, heated to 110 °C, and stirred at this temperature until considered complete (e.g., using TLC or LC/MS analysis). Once complete, the reaction was cooled, treated with excess methanol, and stirred vigorously for 1 hour at room temperature. The slurry was filtered, and the resulting solid cake was washed with methanol. The filtrate was concentrated in vacuo and the resulting solids were resuspended in a solvent mixture of ethyl acetate:isopropyl alcohol = 5:1. The reaction was further treated with saturated sodium carbonate solution and stirred for 10 minutes before separating the layers. The aqueous layer was extracted with the solvent mixture of ethyl acetate:isopropyl alcohol = 5:1 (3 times), and the combined organic materials were washed with brine. The organic substances were further dried over MgSO4, filtered, and the solvent was removed in vacuo. The resulting amidine D was used as is in subsequent steps without further purification.

Stage 4:

Formation of pyrimidone:Amidine D was suspended in ethanol and shaken vigorously at 23 °C to promote complete solvation. The reaction was further treated with sodium 3-ethoxy-2-fluoro-3-oxoprop-1-en-1-olate (e.g., 3 equiv.), and the flask was equipped with a reflux condenser. The reaction was placed in a preheated oil bath maintained at 90 °C and stirred until full consumption of starting material was observed by LC/MS (reaction times were typically 1 h). The contents were cooled to 23 °C, and the reaction mixture was acidified with HCl (e.g., 3 equiv., 1.25 ml solution in EtOH). The mixture was stirred for 30 minutes and most of the solvent was removed in vacuo. The contents were resuspended in ether and water (1:1 mixture), and the resulting suspension was stirred for 20 min. The suspension was filtered under vacuum, and the solid cake was rinsed with additional water and ether and dried under high vacuum overnight. The resulting pyrimidone E was used as is in subsequent steps without further purification.

General procedure B

A solution of amino-type nucleophile (3 equiv.), triethylamine (10 equiv.), and intermediate 1 (1 equiv.) was stirred in dioxane and water (2:1 ratio) at 90 °C until observed by LC/MS the complete consumption of starting material. The solution was diluted with aqueous 1N hydrochloric acid and dichloromethane. The layers were then separated and the aqueous layer was extracted with dichloromethane. The organic substances were combined, dried over magnesium sulfate, filtered, and the solvent was removed in vacuo. Purification produced the desired product.

General procedure C

A mixture of intermediate 2 (this intermediate is described in previously published patent application WO2012/3405 A1; 1 equivalent) and carboxylic acid (1.1 equivalent) in W,W-dimethylformamide was treated with triethylamine (4 equivalents). followed by a solution of 50% ethyl acetate in propylphosphonic anhydride ethyl acetate (T3P, 1.4 equivalents). The reaction was heated to 80 °C for 24 h, after which the reaction was diluted with water and 1N hydrochloric acid solution. The content was extracted with dichloromethane and then ethyl acetate. The combined organic layers were dried over sodium sulfate, filtered, and concentrated in vacuo. Purification produced the desired product.

Synthesis of intermediate product 1

A suspension of 5-fluoro-2-(1-(2-fluorobenzyl)-5-(isoxazol-3-yl)-1H-pyrazol-3-yl)-

-pyrimidin-4-ol (generated by general procedure A, using 1-(isoxazol-3-yl)ethanone in step 1 and 2-fluorobenzylhydrazine in step 2, 11.5 g, 32.4 mmol, 1 equiv .) in phosphoryl trichloride (60.3 ml, 647 mmol, 20 equiv.) at 60 °C for 3 h. The solution was cooled to 23 °C, and poured gradually over the course of 15 min into ice bath water (800 ml) with stirring. After completion of the addition, the contents were stirred for a further 15 min and diluted with dichloromethane (500 mL). The layers were separated and the aqueous layer was extracted with dichloromethane (2 x 200 ml). The organic substances were dried over magnesium sulfate, filtered, and the solvent was removed in vacuo to obtain intermediate product 1 (12.5 g, 103% yield) as a tan solid.

1H NMR (500 MHz, DMSO-cfe) 89.11 (d, 1 H), 9.04 (s, 1 H), 7.71-7.68 (m, 1 H), 7.37-7, 30 (m, 2 H), 7.25-7.20 (m, 1 H), 7.12 (t, 1 H), 6.92 (td, 1 H), 5.95 (s, 2 H) ).

Reference compound I-248

A mixture of intermediate 1 (48 mg, 1 equiv.), (R)-3-methyl-2-((methylamino)methyl)butanoic acid, (99 mg, TFA salt, 3 equiv.), and triethylamine was heated. (0.177 ml, 10 equiv.) at 100 °C as a solution in dioxane/water (2:1) for 20 h, following general procedure B. The content was treated with 3N HCl, and divided between a mixture 1: 1 of dichloromethane and water. The layers were separated and the aqueous layer was treated with a small amount of sodium chloride. The aqueous layer was then extracted with dichloromethane (3 times), and the organic parts were combined and washed with brine. The mixture was dried over MgSO4, filtered and concentrated in vacuo. The crude material was purified by silica gel chromatography with a gradient of 0-10% methanol/dichloromethane providing the desired compound, reference compound I-248 (20 mg, 93%) as an off-white solid.

1H-NMR (500 MHz, MeOD) 88.74 (d, 1 H), 8.09 (d, 1 H), 7.38 (s, 1 H), 7.29-7.23 (m, 1 H), 7.10-7.05 (m, 1 H), 7.02 (td, 1 H), 6.87-6.83 (m, 1 H), 6.83 (d, 1 H) , 5.98-5.89 (m, 2 H), 4.15 (dd, 1 H), 3.81 (dd, 1 H), 3.33 (d, 3 H), 2.72-2 .65 (m, 1 H), 1.94 (dq, 1 H), 1.09 (d, 3 H), 1.01 (d, 3 H).

Reference compound I-250

The title compound was prepared following general procedure B, except that 1-((methylamino)methyl)cyclopropanecarboxylic acid (as the TFA salt) was the amine reagent, the contents were heated at 100 °C for 20 h and the Aqueous layer during processing was treated with sodium chloride. The crude material was purified by silica gel chromatography with a gradient of 0-10% methanol/dichloromethane providing the desired compound, reference compound I-250 (40 mg, 54%) as an off-white solid.

1H-NMR (500 MHz, MeOD) 88.74 (d, 1 H), 8.07 (d, 1 H), 7.36 (s, 1 H), 7.29-7.23 (m, 1 H), 7.11-7.05 (m, 1 H), 7.03 (td, 1 H), 6.88 (d, 1 H), 6.85 (td, 1 H), 5.93 (s, 2 H), 4.14 (s, 2 H), 3.35 (d, 3 H), 1.30-1.25 (m, 2 H), 1.07-1.03 (m , 2H).

Reference compound I-252

The title compound was prepared following general procedure B, except that 2-ethyl-2-((methylamino)methyl)butanoic acid (as the TFA salt) was the amine reagent, the contents were heated to 100 °C for 20 h and the aqueous layer during processing was treated with sodium chloride. The crude material was purified by silica gel chromatography with a gradient of 0-10% methanol/dichloromethane providing the desired compound, reference compound I-252 (33 mg, 39%) as a white solid.

1H-NMR (500 MHz, CD3OD) 88.80 (d, 1 H), 8.25 (d, 1 H), 7.50 (s, 1 H), 7.32-7.26 (m, 1 H), 7.12-7.06 (m, 1 H), 7.04 (t, 1 H), 6.94 (t, 1 H), 6.91 (d, 1 H), 5.97 (s, 2 H), 4.20 (s, 2 H), 3.46 (d, 3 H), 1.86-1.77 (m, 2 H), 1.68 (dq, 2 H) , 0.91 (t, 6 H).

Reference compound I-253

The title compound was prepared following general procedure B, except that (S)-3-methyl-2-((methylamino)methyl)butanoic acid (as the TFA salt) was the amine reagent, the contents were heated at 100 °C for 20 h and the aqueous layer during processing was treated with sodium chloride. The crude material was purified by silica gel chromatography with a gradient of 0-10% methanol/dichloromethane providing the desired compound, reference compound I-253 (26 mg, 64%) as a white solid.

1H-NMR (500 MHz, CD3OD) 88.74 (d, 1 H), 8.08 (d, 1 H), 7.37 (s, 1 H), 7.28-7.22 (m, 1 H), 7.10-7.05 (m, 1 H), 7.02 (t, 1 H), 6.84 (t, 1 H), 6.82 (d, 1 H), 5.97 -5.88 (m, 2 H), 4.15 (dd, 1 H), 3.79 (dd, 1 H), 3.32 (d, 3 H), 2.70-2.64 (m , 1 H), 1.93 (dq, 1 H), 1.08 (d, 3 H), 1.01 (d, 3 H).

Reference compound I-260

The title compound was prepared following general procedure B, except that 4-benzylpiperidine-4-carboxylic acid was the amine reagent, the content was heated at 100 °C for 20 h, and the aqueous layer during processing was treated with chloride of sodium. The crude material was purified by silica gel chromatography with a gradient of 0-10% methanol/dichloromethane providing the desired reference compound I-260 (26 mg, 64%) as a white solid.

1H-NMR (500 MHz, CD3OD) 88.74 (d, 1 H), 8.11 (d, 1 H), 7.41 (s, 1 H), 7.29-7.22 (m, 3 H), 7.22-7.15 (m, 3 H), 7.11 7.06 (m, 1 H), 7.05-7.00 (m, 1 H), 6.91 (d, 1 H), 6.84-6.79 (m, 1 H), 5.96 (s, 2 H), 4.57 (d, 2 H), 3.29-3.23 (m, 2 H ), 2.90 (s, 2 H), 2.19 (d, 2 H), 1.68 - 1.61 (m, 2 H).

Reference compound I-262

The title compound was prepared following general procedure B, except that ethyl 2-methylpiperidine-2-carboxylate was the amine reagent, the content was heated at 100 °C for 19 h, and the aqueous layer during processing was treated with sodium chloride. The crude material was purified by reverse phase HPLC using a gradient of 5-75% acetonitrile/water providing the desired compound, reference compound I-262 (1.1 mg, 8%) as a white solid.

1H-NMR (500 MHz, CD3OD) 88.82 (d, 1 H), 8.33 (d, 1 H), 7.47 (s, 1 H), 7.32-7.26 (m, 1 H), 7.12-7.07 (m, 1 H), 7.05 (t, 1 H), 6.92 (t, 1 H), 6.88 (d, 1 H), 6.03 -5.95 (m, 2 H), 4.32-4.24 (m, 1 H), 3.63 (dt, 1 H), 2.14 (ddd, 1 H), 2.01-1 .79 (m, 5 H), 1.76 (s, 3 H).

Reference compound I-265

The title compound was prepared following general procedure B, except that 3-phenylpyrrolidine-3-carboxylic acid was the amine reagent, the content was heated at 100 °C for 24 h, and the aqueous layer during processing was treated with chloride of sodium. The crude material was purified by silica gel chromatography with a gradient of 0-10% methanol/dichloromethane providing the desired compound, reference compound I-265 (29 mg, 45%) as an off-white solid.

1H-NMR (500 MHz, CD3OD) 88.74 (d, 1 H), 8.11 (d, 1 H), 7.51-7.44 (m, 3 H), 7.40-7.36 (m, 2 H), 7.32-7.23 (m, 2 H), 7.12-7.06 (m, 1 H), 7.03 (t, 1 H), 6.92 (s , 1 H), 6.81 (t, 1 H), 5.96 (s, 2 H), 4.03-3.96 (m, 1 H), 3.91 (d, 1 H), 3 .87 (br. s., 1 H), 3.07-3.00 (m, 1 H), 2.41-2.32 (m, 1 H).

REFERENCE COMPOSITION I-267

The title compound was prepared following general procedure B, except that 3,3-dimethylpiperidine-2-carboxylic acid (as the HCl salt) was the amine reagent, the contents were heated at 100 °C for 18 h and the Aqueous layer during processing was treated with sodium chloride. The crude material was purified by silica gel chromatography with a gradient of 0-10% methanol/dichloromethane providing the desired compound, reference compound I-267 (15 mg, 17%) as a white solid.

1H-NMR (500 MHz, CD3OD) 88.81 (d, 1 H), 8.35 (d, 1 H), 7.57 (s, 1 H), 7.32-7.26 (m, 1 H), 7.12-7.07 (m, 1 H), 7.04 (t, 1 H), 6.94-6.90 (m, 2 H), 5.99 (s, 2 H) , 4.99 (s, 1 H), 4.62 (d, 1 H), 3.86 (td, 1 H), 2.07-1.96 (m, 1 H), 1.95-1 .87 (m, 1 H), 1.81-1.75 (m, 1 H), 1.50 (d, 1 H), 1.22 (s, 3 H), 1.17 (s, 3 H).

Reference compound I-269

The title compound was prepared following general procedure B, except that 3-aminobicyclo[1.1.1]pentane-1-carboxylic acid (as the TFA salt) was the amine reagent, the contents were heated to 100 °C for 18 h and the aqueous layer during processing was treated with sodium chloride.

The crude material was purified by silica gel chromatography with a gradient of 0-10% methanol/dichloromethane providing the desired compound, reference compound I-269 (11 mg, 16%) as a white solid.

1H-NMR (500 MHz, CD3OD 88.76 (d, 1 H), 8.08 (d, 1 H), 7.36 (s, 1 H), 7.30-7.23 (m, 1 H ), 7.12-7.06 (m, 1 H), 7.04 (t, 1 H), 6.96 (d, 1 H), 6.91 (t, 1 H), 5.94 ( s, 2 H), 2.53 (s, 6 H).

Reference compound I-80

The title compound was prepared following general procedure B, except that L-phenylalanine was the amine reagent and the content was heated to 90 °C for 48 h as a solution in THF/water (2:1). The contents were concentrated in vacuo and the crude material was purified by reverse phase HPLC using a gradient of 5-75% acetonitrile/water providing the desired product, reference compound I-80 (1.3 mg, 4%) as a white solid.

1H-MRI (500 MHz, CD3ODMeOD) 88.81 (s, 1 H), 8.20 (d, 1 H), 7.51-7.48 (m, 1 H), 7.34- 7.26 (m, 3 H), 7.22 (t, 2 H), 7.17-7.03 (m, 3 H), 6.96 (s, 1 H), 6.90 (t, 1 H) , 6.00 (s, 2 H), 5.36-5.29 (m, 1 H), 3.48 (d, 1 H), 3.24-3.18 (m, 1 H).

Reference compound I-81

The title compound was prepared following general procedure B, except that L-tryptophan was the amine reagent and the content was heated to 90 °C for 48 h as a solution in THF/water (2:1). The contents were concentrated in vacuo and then purified by reverse phase HPLC using a gradient of 5-75% acetonitrile/water to provide the desired compound, reference compound I-81 (7.3 mg, 18%) as a solid. brown.

1H-NMR (500 MHz, CD3OD) 88.86-8.83 (m, 1 H), 8.16 (d, 1 H), 7.69 (d, 1 H), 7.33-7.27 (m, 1 H), 7.16 (d, 1 H), 7.13 7.04 (m, 4 H), 7.01-6.96 (m, 1 H), 6.95-6, 88 (m, 3 H), 5.96 (s, 2 H), 5.51 (dd, 1 H), 3.74-3.67 (m, 1 H), 3.30-3.25 ( m, 1H).

Reference compound I-85

The title compound was prepared following general procedure B, except that 1-aminocyclopropanecarboxylic acid was the amine reagent and the content was heated at 90 °C for 48 h as a solution in THF/water (2:1). The contents were concentrated in vacuo and the residue was purified by reverse phase HPLC using a gradient of 5-95% acetonitrile/water providing the desired compound, reference compound I-85 (7.3 mg, 18%) as a transparent oil.

1H-NMR (500 MHz, CD3OD) 88.83 (d, 1 H), 8.38 (d, 1 H), 7.47 (s, 1 H), 7.34-7.28 (m, 1 H), 7.13-7.04 (m, 2 H), 6.99 6.95 (m, 2 H), 6.02 (s, 2 H), 1.84-1.79 (m, 2H), 1.43-1.38 (m, 2H).

Reference compound I-93

The title compound was prepared following general procedure B, except that (3-aminooxetan-3-yl)methanol was the amine reagent and the contents were heated at 170 °C for 15 min in the microwave as a solution in THF. /water (2:1). The contents were concentrated in vacuo and the residue was purified by reverse phase HPLC using a gradient of 5-75% acetonitrile/water providing the desired compound, reference compound I-93 (0.6 mg, 4%) as a transparent oil.

<1>H -R M N (500 M H z, C D<3>O D ) 8 8 .85 (d, 1 H), 8 .55 (s, 1 H), 7 .69 (s, 1 H), 7 , 32 - 7 .37 (m, 1 H), 7 .09 - 7 .17 (m, 3 H), 6 .97 (d, 1 H), 6.01 (s, 2 H), 5 .00 ( s, 2 H), 3 .76 (q, 4 H).

I-102 REFERENCE COMPOSITION

The title compound was prepared following general procedure B, except that methyl 2-amino-2-(oxetan-3-yl) acetate was the amine reagent and the contents were heated at 100 °C for 42 h as a solution in THF/water (2:1). The contents were concentrated in vacuo and the residue was purified by reverse phase HPLC using a gradient of 5-75% acetonitrile/water providing the desired compound, reference compound I-102 (0.6 mg, 2%) as a transparent oil.

1H-MRI (500 MHz, CD3OD) 88.80 (d, 1 H), 8.30 (d, 1 H), 7.50 (s, 1 H), 7.32-7.27 (m, 1 H), 7.12-7.03 (m, 2 H), 6.92 (t, 1 H), 6.89 (d, 1 H), 5.99 (s, 2 H), 5.23 (d, 1 H), 4.65 (t, 1 H), 4.31 (t, 1 H), 3.83-3.74 (m, 2 H), 3.02 (dtd, 1 H) .

Reference compound I-109

The title compound was prepared following general procedure B, except that no amine reagent was used, DBU was used instead of triethylamine, and the contents were heated at 100 °C for 18 h as a solution in THF/water (2 :1). The contents were concentrated in vacuo and the residue was purified by reverse phase HPLC using a gradient of 5-75% acetonitrile/water to provide the desired compound, reference compound I-109 (7 mg, 35%) as a clear oil. .

1H-NMR (500 MHz, CD3OD) 88.84 (d, 1 H), 8.26 (d, 1 H), 7.67 (s, 1 H), 7.25-7.28 (m, 1 H), 7.14-7.05 (m, 2 H), 7.02 (d, 1 H), 7.01- 6.97 (m, 1 H), 6.03 (s, 2 H) , 3.79 (t, 2 H), 3.56-3.47 (m, 4 H), 2.56-2.50 (m, 2 H), 1.99 (quintuplet, 2 H), 1 .80-1.73 (m, 2 H), 1.72 -1.61 (m, 4 H).

Reference compound I-108

The title compound was prepared following general procedure B, except that D-tryptophan was the amine reagent and the content was heated to 100 °C for 18 h as a solution in THF/water (2:1). The contents were treated with 3N HCl solution, the solvent was removed in vacuo and the resulting solid was washed with H2O, then purified by reverse phase HPLC using a gradient of 5-75% acetonitrile/water providing the desired compound, the reference compound I-108 (3.5 mg, 16%) as a clear oil.

1H-NMR (500 MHz, CD3OD) 88.85 (d, 1 H), 8.16 (d, 1 H), 7.69 (d, 1 H), 7.33-7.27 (m, 1 H), 7.17 (d, 1 H), 7.13-7.05 (m, 4 H), 7.01-6.96 (m, 1 H), 6.95-6.89 (m , 3 H), 5.97 (s, 2 H), 5.50 (dd, 1 H), 3.70 (dd, 1 H), 3.28 (d, 1 H).

Reference compound I-116

The title compound was prepared following general procedure B, except that D-phenylalanine was the amine reagent and the content was heated to 100 °C for 18 h as a solution in THF/water (2:1). The contents were treated with 3N HCl solution, the solvent was removed in vacuo and the resulting residue was purified by reverse phase HPCL with a gradient of 5-75% acetonitrile/water providing the desired compound, reference compound I-116. (25 mg, 61%) as a solid.

1H-MRI (500 MHz, CD3OD, MeOD) 88.77 (s, 1 H), 8.13 (d, 1 H), 7.43 (s, 1 H), 7.31 (d, 2 H) , 7.28-7.18 (m, 3 H), 7.16 7.11 (m, 1 H), 7.09-7.03 (m, 1 H), 7.01 (t, 1 H) ), 6.91 (s, 1 H), 6.85 (t, 1 H), 5.94 (s, 2 H), 5.26 (dd, 1 H), 3.45 (dd, 1 H ), 3.19 (dd, 1 H).

Reference compound I-117

The title compound was prepared following general procedure B, except that L-phenylglycine was the amine reagent and the content was heated to 100 °C for 18 h as a solution in THF/water (2:1). The contents were treated with 3N HCl solution, the solvent was removed in vacuo and the resulting solid was purified by reverse phase HPCL with a gradient of 5-75% acetonitrile/water providing the desired compound, reference compound I-117. (26 mg, 63%) as a solid.

1H-NMR (500 MHz, CD3OD) 88.81 (s, 1 H), 8.29 (d, 1 H), 7.61 (d, 2 H), 7.52 (s, 1 H), 7 .46-7.36 (m, 3 H), 7.27 (q, 1 H), 7.10-7.05 (m, 1 H), 7.03 (t, 1 H), 6.95 -6.90 (m, 2 H), 6.02 (s, 1 H), 5.97 (s, 2 H).

Reference compound I-118

The title compound was prepared following general procedure B, except that D-phenylglycine was the amine reagent and the content was heated to 100 °C for 18 h as a solution in THF/water (2:1). The contents were treated with 3N HCl solution, the solvent was removed in vacuo and the resulting solid was purified by reverse phase HPCL with a gradient of 5-75% acetonitrile/water providing the desired compound, reference compound I-118. (22 mg, 53%) as a solid.

<1>H -R M N (500 M Hz, C D<3>O D ) 8 8.81 (s, 1 H), 8 .30 (d, 1 H), 7 .60 (d, 2 H), 7 , 53 (s, 1 H), 7 .46 -7 .37 (m, 3 H), 7 .28 (q, 1 H), 7 .11 -7 .06 (m, 1 H), 7 .04 ( t, 1 H), 6 .96-6 .91 (m, 2 H), 6 .02 (s, 1 H), 5 .99 (s, 2 H).

REFERENCE COMPOSITION I-142

The title compound was prepared following general procedure B, except that N-methyl-phenylglycine was the amine reagent and the content was heated at 100 °C for 18 h as a solution in THF/water (2:1). The contents were treated with 3N HCl solution, the solvent was removed in vacuo and the resulting solid was purified by reverse phase HPCL with a gradient of 5-75% acetonitrile/water providing the desired compound (15 mg, 52%) as a solid.

1H-NMR (500 MHz, MeOD) 88.80 (d, 1 H), 8.45-8.39 (m, 1 H), 7.58-7.55 (m, 1 H), 7.50 -7.44 (m, 5 H), 7.34-7.27 (m,

1 H), 7.14-7.04 (m, 2 H), 7.00-6.94 (m, 1 H), 6.90 (d, 1 H), 6.61-6.55 ( m, 1 H), 6.02 (s, 2 H), 3.25-3.20 (m, 3 H).

Reference compound I-120

The title compound was prepared following general procedure B, except that 1-(aminomethyl)cyclopropanecarboxylic acid was the amine reagent, the content was heated at 100 °C for 22 h, and the aqueous layer during processing was treated with sodium chloride. sodium. The crude material was purified by silica gel chromatography with a gradient of 0-10% methanol/dichloromethane providing the desired compound, reference compound I-120 (20 mg, 42%) as a white solid.

1H-NMR (500 MHz, CD3OD) 88.75 (d, 1 H), 8.05 (d, 1 H), 7.39 (s, 1 H), 7.30-7.24 (m, 1 H), 7.12-7.06 (m, 1 H), 7.03

(t, 1 H), 6.89 (d, 1 H), 6.84 (t, 1 H), 5.95 (s, 2 H), 3.88 (s, 2 H), 1.25 -1.20 (m, 2 H), 1.15-1.10 (m, 2 H).

Reference compound I-207

The title compound was prepared following general procedure B, except that N-methyl-L-alanine was the amine reagent, the content was heated to 100 °C for 22 h, and the aqueous layer during preparation was treated with sodium chloride. The crude material was purified by silica gel chromatography with a gradient of 0-10% methanol/dichloromethane providing the desired compound, reference compound I-207 (20 mg, 57%) as a white solid.

1H-NMR (500 MHz, CD3OD) 88.74 (d, 1 H), 8.16 (d, 1 H), 7.40 (s, 1 H), 7.29-7.23 (m, 1 H), 7.11-7.05 (m, 1 H), 7.02

(t, 1 H), 6.87 (d, 1 H), 6.82 (t, 1 H), 5.94 (s, 2 H), 5.10 (q, 1 H), 3.33 (d, 3 H), 1.59 (d, 3 H).

Reference compound I-217

The title compound was prepared following general procedure B, except that 2-(aminomethyl)-2-ethylbutanoic acid was the amine reagent, the content was heated to 100 °C for 22 h, and the aqueous layer during processing It was treated with sodium chloride. The crude material was purified by silica gel chromatography with a gradient of 0-10% methanol/dichloromethane providing the desired compound, reference compound I-217 (20 mg, 50%) as a clear solid.

1H-NMR (500 MHz, CD3OD) 88.76-8.72 (m, 1 H), 8.07-8.03 (m, 1 H), 7.42-7.39 (m, 1 H) , 7.29-7.22 (m, 1 H), 7.11

7.04 (m, 1 H), 7.02 (t, 1 H), 6.89-6.81 (m, 2 H), 5.94 (s, 2 H), 3.91 (s, 2 H), 1.68 (q, 4 H), 0.98-0.90 (t, 6 H).

Reference compound I-224 and compound I-225

The title compounds were prepared following general procedure B, except that 2-amino-5,5,5-trifluoro-4-methylpentanoic acid was the amine reagent, the contents were heated at 100 °C for 18 h and the layer aqueous during processing was treated with sodium chloride. The crude material was purified by reverse phase HPLC using a gradient of 5-75% acetonitrile/water providing the desired diastereomers, compound I-224

(3.3 mg, 7%, first eluant in LCMS) as a white solid and the reference compound I-225 (2 mg, 5%, second eluant in LCMS) as a white solid.

1H-NMR for Reference Compound I-224 (500 MHz, CD3OD) 8 8.75 (d, 1 H), 8.15 (d, 1 H), 7.38 7.24 (m, 1 H) , 7.11-7.06 (m, 1 H), 7.03 (t, 1 H), 6.86 (d, 1 H), 6.83 (t, 1 H), 5.95 (s , 2H), 4.94 (t, 1H), 2.60 (dd, 1H),

2.45-2.38 (m, 1 H), 1.96-1.89 (m, 1 H), 1.25 (d, 3 H).

1H-NMR for Reference Compound I-225 (500 MHz, CD3OD) 8 8.81 (d, 1 H), 8.34 (d, 1 H), 7.58 7.27 (m, 1 H) , 7.13-7.08 (m, 1 H), 7.06 (t, 1 H), 6.99-6.92 (m, 2 H), 6.01 (s, 2 H), 5 .26 (dd, 1 H), 2.53-2.42 (m, 2.42-2.33 (m, 1 H), 2.13 (ddd, 1 H), 1.24 (d, 3 H).

Reference compound I-226

The title compound was prepared following general procedure B, except that 2-amino-3-fluoro-3-methylbutanoic acid was the amine reagent, the content was heated at 100 °C for 20 h, and the aqueous layer during processing It was treated with sodium chloride. The crude material was purified by silica gel chromatography with a gradient of 0-10% methanol/dichloromethane providing the desired compound, reference compound I-226 (11 mg, 42%) as a white solid.

<1>H -R M N (500 M Hz, C D<3>O D ) 8 8 .75 (d, 1 H), 8 .16 (d, 1 H), 7 .44 (s, 1 H), 7 , 30 -7 .22 (m, 1 H), 7 .11 -7 .06 (m, 1 H), 7 .02

(t, 1 H), 6 .90 (d, 1 H), 6.81 (t, 1 H), 5 .95 (s, 2 H), 5 .13 (d, 1 H), 1 .65 -1 .58 (m, 3 H), 1 .58 -1 .51 (m , 3 H).

I-227 REFERENCE COMPOSITION

The title compound was prepared following general procedure B, except that (S)-2-amino-2-hydropropylacetic acid was the amine reagent, the content was heated at 100 °C for 20 h, and the aqueous layer during processing It was treated with sodium chloride. The crude material was purified by silica gel chromatography with a gradient of 0-10% methanol/dichloromethane providing the desired compound, reference compound I-227 (21 mg, 86%) as a white solid.

1H-NMR (500 MHz, MeOD) 88.74 (d, 1 H), 8.10 (d, 1 H), 7.37 (s, 1 H), 7.28-7.22 (m, 1 H), 7.11-7.05 (m, 1 H), 7.02 (t, 1 H), 6.85 (d, 1 H), 6.82 (t, 1 H), 5.93 (s, 2 H), 3.96 (d, 1 H), 1.38-1.28 (m, 1 H), 0.75-0.64 (m, 3 H), 0.53-0 .47 (m, 1 H).

Reference compound I-239

The title compound was prepared following general procedure B, except that (S)-N-methyl--2-amino-2-cyclopropylacetic acid was the amine reagent, the contents were heated at 100 °C for 20 h and the Aqueous layer during processing was treated with sodium chloride. The crude material was purified by silica gel chromatography with a gradient of 0-10% methanol/dichloromethane providing the desired compound, reference compound I-239 (4 mg, 20%) as a white solid.

1H-NMR (500 MHz, CD3OD) 88.75 (d, 1 H), 8.16 (d, 1 H), 7.39 (s, 1 H), 7.26 (ddd, 1 H), 7 .08 (ddd, 1 H), 7.04-7.00 (m, 1 H), 6.86 (d, 1 H), 6.82 (td, 1 H), 5.94 (s, 2 H), 4.19 (d, 1 H), 3.48 (d, 3 H), 1.53-1.44 (m, 1 H), 0.91-0.83 (m, 1 H) , 0.76-0.64 (m, 2 H), 0.44 (dq, 1 H).

Reference compound I-240

The title compound was prepared following general procedure B, except that (R)-2-amino-2-cyclopropylacetic acid was the amine reagent, the content was heated to 100 °C for 2 h, and the aqueous layer during processing It was treated with sodium chloride. The crude material was purified by silica gel chromatography with a gradient of 0-10% methanol/dichloromethane providing the desired compound, reference compound I-240 (46 mg, 93%) as a white solid.

1H-NMR (500 MHz, CD3OD) 88.74 (d, 1 H), 8.10 (d, 1 H), 7.36 (s, 1 H), 7.28-7.22 (m, 1 H), 7.07 (ddd, 1 H), 7.01 (td, 1 H), 6.84 (d, 1 H), 6.81 (td, 1 H), 5.93 (s, 2 H), 3.96 (d, 1 H), 1.38-1.30 (m, 1 H), 0.74-0.65 (m, 3 H), 0.52-0.47 (m , 1 HOUR).

Reference compound I-241

The title compound was prepared following general procedure B, except that (R)-N-methyl-2-amino-2-cyclopropylacetic acid (as TFA salt) was the amine reagent, the contents were heated to 100° C for 20 h and the aqueous layer during processing was treated with sodium chloride. The crude material was purified by silica gel chromatography with a gradient of 0-10% methanol/dichloromethane providing the desired compound, reference compound I-241 (20 mg, 93%) as a white solid.

1H-NMR (500 MHz, CD3OD) 88.74 (d, 1 H), 8.15 (d, 1 H), 7.38 (s, 1 H), 7.28-7.22 (m, 1 H), 7.10-7.04 (m, 1 H), 7.04 6.99 (m, 1 H), 6.85 (d, 1 H), 6.82 (t, 1 H), 5.93 (s, 2 H), 4.18 (d, 1 H), 3.48 (d, 3 H), 1.53-1.44 (m, 1 H), 0.91-0, 82 (m, 1 H), 0.76-0.64 (m, 2 H), 0.48-0.41 (m, 1 H).

Reference compound I-90

The title compound was prepared following general procedure B, except that (S)-indoline-2-carboxylic acid was the amine reagent (1 equiv.), and the contents were heated at 90 °C for 12 h as a solution in THF/water (1:1), followed by heating at 125 °C for 15 min in the microwave. The contents were extracted with ethyl acetate during processing. The crude material was purified by reverse phase HPLC using 5 to 95% acetonitrile in water with addition of 0.1% trifluoacetic acid to provide the desired compound, reference compound I-90 (3.9 mg, 15% yield) as a whitish solid.

1H-NMR (500 MHz, DMSO-de) 8 (ppm): 9.10 - 9.21 (d, 1 H), 8.61 - 8.75 (m, 1 H), 8.47 - 8, 57 (d, 1 H), 7.49 - 7.58 (s, 1 H), 7.33 - 7.41 (m, 1 H), 7.22 - 7.33 (m, 4 H), 7.10 - 7.20 (m, 2 H), 6.98 - 7.10 (m, 1 H), 5.95 (s, 2 H), 5.39 - 5.53 (m, 1 H) ), 3.64 - 3.74 (dd, 1 H), 3.20 - 3.32 (dd, 2 H).

Reference compound I-91

The title compound was prepared following general procedure B, except that (R)-indoline-2-carboxylic acid was the amine reagent (1 equiv.), and the contents were heated at 90 °C for 12 h as a solution in THF/Water (1:1), followed by heating at 125 °C for 15 min in the microwave. The content was extracted with ethyl acetate during work. The crude material was purified by reverse phase HPLC using a gradient of 5-95% acetonitrile in water (in 0.1% TFA) providing the desired compound, reference compound I-91 obtained following the usual procedure (1, 9mg, 7%).

1H NMR (500 MHz, CD3CN) 5 (ppm): 8.68 - 8.75 (d, 1 H), 8.35 - 8.49 (m, 2 H), 7.42 -7.49 (m , 1 H), 7.27 - 7.41 (m, 3 H), 7.05 - 7.24 (m, 4 H), 6.91 - 6.96 (m, 1 H), 5.97 (s, 2 H), 5.38 - 5.48 (m, 1 H), 3.65 - 3.79 (dd, 1 H), 3.31 - 3.44 (dd, 1 H).

Reference compound I-114

Purification was achieved by reverse phase HPLC using 5-75% acetonitrile in water for 30 minutes (with addition of 0.1% trifluoacetic acid) providing the desired compound (1.6 mg, 4% yield) as an oil. transparent. Only the last diastereomer (reference compound I-114) was purified from this reaction mixture.

1H NMR (500 MHz, 500 MHz, CD3CN) 5 (ppm): 8.85 (s, 1 H), 8.33 (d, 1 H), 7.40 - 7.48 (m, 1 H), 7.28 - 7.38 (m, 1 H), 7.04 - 7.19 (m, 2 H), 6.90 - 7.00 (m, 2 H), 6.03 (s, 2 H) ), 3.13 - 3.17 (m, 1 H), 2.47 - 2.59 (m, 1 H), 2.36 - 2.42 (m, 1 H), 2.03-2, 17 (m, 1 H), 1.77 - 1.85 (m, 1 H), 1.65 - 1.74 (m, 2 H), 1.49 - 1.60 (m, 2 H), 1.38- 1.47 (m, 1 H).

Reference compound I-107

The title compound was prepared following general procedure B, except that (1S,2S,5R)-3-azabicyclo[3.1.0]hexane-2-carboxylic acid was the amine reagent (1 equiv.), 3 equivalents of triethylamine, and the content was heated at 70 °C for 14 h as a solution in THF/water (10:1). The content was extracted with ethyl acetate during processing, dried, filtered and concentrated, obtaining the desired compound. Reference compound I-107 (38.3 mg, 100% yield) was obtained as a tan solid. No purification was needed for this compound.

1H NMR (500 MHz, CD3OD) 5 ppm: 8.79 (s, 1 H), 8.23 (d, 1 H), 7.36-7.46 (br. s, 1 H), 7.25 -7.31 (m, 1 H), 7 .06 -7 .12 (m, 1 H), 7.01 - 7.06 (m, 1 H), 6.83 - 6.90 (m, 2 H, 2 overlapping movements), 5.96 (s, 2 H), 4.18 (dd, 1 H), 4.02 -4.08 (m, 1 H), 1.93 - 2.02 (m , 1 H), 0.83 - 0.93 (m, 4 H).

Reference compound I-129

Purification was achieved by silica gel chromatography using 1 to 10% methanol in dichloromethane for 30 minutes providing compound I-129 (21.7 mg, 57% yield) as a white solid.

1H NMR (400 MHz, CDCh) 5 (ppm): 8.45 - 8.57 (m, 2 H, 2 isochronous shifts) 7.40 - 7.48 (m, 3 H), 7.24 -7, 40 (m, 1 H), 6.93 -7.09 (m, 2 H), 6.58-6.68 (m, 1 H), 5.90 (s, 2 H), 3.74- 3.90 (m, 2 H), 1.99 -2.20 (m, 2 H),1.70-1.89 (m, 4 H), 1.55- 1.69 (m, 2 H) ).

Reference compound I-124

The title compound was prepared following general procedure B, except that 4-methylpiperidine-4-carboxylic acid (as the HCl salt) was the amine reagent (1.1 equiv.), 4 equivalents of triethylamine were used, and the content was heated at 80 °C for 18 h as a solution in THF/water (10:1). The content was extracted with ethyl acetate during work. The crude material was purified by silica gel chromatography with a gradient of 1-10% methanol/dichloromethane over 30 minutes providing the desired compound, reference compound I-124 as an off-white solid (36.1 mg, 95% yield). .

1H NMR (400 MHz, CDCh) 5 (ppm): 8.49 (s, 1 H), 8.16 - 8.28 (d, 1 H), 7.35 -7.44 (m, 1 H) , 7.17 - 7.26 (m, 1 H), 6.95 -7.10 (m, 2 H), 6.87 (m, 1 H), 6.62 (s, 1 H), 6 .00 (s, 2 H), 4.34 - 4.48 (m, 1 H), 3.36 - 3.48 (m, 1 H), 2.36 - 2.41 (m, 1 H) , 1.58 - 1.68 (m, 1 H), 1.34 (s, 3 H), 0.71 -0.81 (m, 4 H).

Reference compound I-143

The title compound was prepared following general procedure B, except that 3-methylpyrrolidine-3-carboxylic acid was the amine reagent (1.05 equiv.), 4 equivalents of triethylamine were used, and the contents were heated to 80 °C for 4 h as a solution in THF/water (10:1). The content was extracted with ethyl acetate during work. The crude material was purified by silica gel chromatography using a gradient of 1-10% methanol/dichloromethane over 30 minutes providing the desired compound, reference compound I-143 as a white solid (18.9 mg, 48% yield ).

1H NMR (500 MHz, CDCh) 5 ppm: 8.45 (s, 1 H), 8.12 - 8.19 (d, 1 H), 7.30 (s, 1 H), 7.27 (s , 1 H), 7.14 - 7.22 (m, 1 H), 6.98 - 7.05 (m, 1 H), 6.93 - 6.98 (m, 1 H), 6.80 - 6.87 (m, 1 H), 6.57 (d, 1 H), 5.96 (s, 2 H), 4.24 - 4.36 (m, 1 H), 3.84 -4 .00 (m, 2 H), 3.59 - 3.70 (m, 1 H), 2.45 - 2.58 (m, 1 H), 1.84 - 2.00 (m, 1 H) , 1.47 (s, 3 H).

Reference compound I-152

The title compound was prepared following general procedure B, except that 4,4-dimethyl-pyrrolidine-3-carboxylic acid was the amine reagent (1.05 equiv.), 4 equivalents of triethylamine were used, and the content was heated to 90 °C for 14 h as a solution in THF/water (10:1). The content was extracted with ethyl acetate during work. The crude material was purified by silica gel chromatography with a gradient of 1-7% methanol/dichloromethane over 30 minutes providing the desired compound, reference compound I-152 as an off-white solid (14.3 mg, 37% yield ).

1H NMR (400 MHz, CDCh) 8 (ppm): 8.45 (s, 1 H), 8.05 - 8.20 (d, 1 H), 7.29 -7.34 (m, 1 H) , 7.14 - 7.25 (m, 1 H), 6.91 -7.08 (m, 2 H), 6.79 - 6.87 (m, 1 H), 6.56 - 6.63 (m, 1 H), 5.96 (s, 2 H), 4.01 - 4.23 (m, 2 H), 3.71 - 3.87 (dd, 1 H), 3.53 -3 .65 (dd, 1 H), 2 .85-2.97 (m, 1 H), 1.34 (s, 3 H), 1.15 (s, 3 H).

Reference compound I-186

The title compound was prepared following general procedure B, except that 4-phenylpiperidine-4-carboxylic acid (as the HCl salt) was the amine reagent (1.05 equiv.), 4 equivalents of triethylamine were used, and the content was heated at 70 °C for 24 h as a solution in THF/water (10:1). The content was extracted with ethyl acetate during work. The crude material was purified by silica gel chromatography using a gradient of 4-7% methanol/dichloromethane over 40 minutes providing the desired compound, reference compound I-186 as a white solid (22.3 mg, 51% yield ).

1H NMR (400 MHz, CDCh) 8 (ppm): 8.46 (s, 1 H), 8.19 (d, 1 H), 7.44 -7.49 (m, 2 H, 2 overlapping movements) , 7.36 - 7.41 (m, 2 H), 7.29 - 7.34 (m, 2 H), 7.16 - 7.22 (m, 2 H), 6.99 - 7.05 (m, 1 H), 6.93 - 6.98 (m, 1 H), 6.81 - 6.86 (m, 1 H), 6.59 (m, 1 H), 5.97 (s , 2 H), 4.50 - 4.58 (m, 2 H), 3.42 - 3.50 (m, 2 H), 2.69 - 2.75 (m, 2 H), 2.07 -2.14 (m, 2H).

Reference compound I-194

This compound was prepared following general procedure B described above, except that 4-(aminomethyl)tetrahydro-2 H-pyran-carboxylic acid was the amine reagent (1.05 equiv.), 4 equivalents of triethylamine were used, and The contents were heated at 70 °C for 6 h as a solution in THF/water (10:1), followed by heating at 90 °C for 12 h. The contents were extracted with ethyl acetate during preparation and purification was achieved by silica gel chromatography using 4 to 7% methanol in dichloromethane for 40 minutes providing the desired compound, reference compound I-194 ((26, 8 mg, 66% yield) as a white solid

1H NMR (500 MHz, CDCla) 8 (ppm): 8.46 (s, 1 H), 8.10 (d, 1 H), 7.28 (s, 1 H), 7.18 - 7.24 (m, 1 H), 6.94 - 7.07 (m, 3 H), 6.58 (d, 1 H), 5.95 (s, 2 H), 5.50 - 5.57 (m , 1 H), 3.86 - 3.94 (m, 2 H), 3.79 - 3.85 (m, 2 H), 3.51 - 3.60 (m, 2 H), 2.12 -2.20 (m, 2 H), 1.53- 1.62 (m, 2 H).

Reference compound I-228

The title compound was prepared in 4 steps:

Stage 1: 1-((4-methylphenylsulfonamido)methyl)cyclopentanecarboxylic acid

A suspension of 1-(aminomethyl)cyclopentanecarboxylic acid (316 mg, 1.0 equiv), p-toluenesulfonyl chloride (505 mg, 1.2 equiv), and 1 M sodium hydroxide aqueous solution (6.62 mL, 3.0 equiv.) in water (10 ml) was heated to 90 °C for 1 h, after which the reaction mixture was cooled to 0 °C and acidified by adding an aqueous solution of hydrochloric acid to the 3M. The resulting white precipitate was filtered, then washed successively with water and ethanol to provide 1-((4-methylphenylsulfonamido)methyl)cyclopentanecarboxylic acid (383 mg, 58% yield) as a white solid.

1H-NMR (500 MHz, DMSO-de) 8 (ppm): 12.14 - 12.38 (s, 1H), 7.64 - 7.75 (d, 2H), 7.47 - 7.56 ( t, 1H), 7.33 - 7.45 (d, 2H), 2.79-2.90 (d, 2H), 2.38 (s, 3H), 1.81 -1.95 (m, 2H), 1.47 -1.65 (m, 6H).

Stage 2: 1-((N,4-dimethylphenylsulfonamido)methyl)cyclopentanecarboxylic acid

A solution of 1-((4-methylphenylsulfonamido)methyl)cyclopentanecarboxylic acid (383 mg, 1.0 equiv.), iodomethane (0.254 ml, 3.15 equiv.) and 1 M aqueous sodium hydroxide solution (5.15 ml, 4.0 equiv.) in water (5 ml) was heated at 75 °C for 1.5 hours, after which analysis by LC/MS indicated that the reaction was complete. The reaction mixture was cooled to room temperature, washed with dichloromethane (3 x 30 ml), acidified by adding 3 M aqueous hydrochloric acid solution, extracted with diethyl ether (3 x 30 ml), dried ( sodium sulfate), filtered and concentrated to provide 1-((N,4-dimethylphenylsulfonamido)methyl)cidopentanecarboxylic acid (343 mg, 86% yield) as a golden yellow solid. It was not necessary to purify.

1H NMR (500 MHz, CDCla) 8 (ppm): 7.59 - 7.73 (d, 2H), 7.30 - 7.41 (d, 2H), 3.24 -3.39 (s, 2H) ), 2.71 (s, 3H), 2.45 (s, 3H), 2.06 - 2.22 (m, 2H), 1.69 - 1.88 (m, 6H).

Step 3: 1-((methylamino)methyl)cyclopentanecarboxylic acid hydrobromide

A solution of 1-((N,4-dimethylphenylsulfonamido)methyl)cyclopentanecarboxylic acid (343 mg, 1.0 equiv.) was heated in a solution of glacial acetic acid to 33% hydrogen bromide (6.0 ml, 30 equiv.) for 2 hours 75 °C. The reaction was then cooled to room temperature, diluted in water (10 ml) and washed with diethyl ether (3 x 40 ml). The aqueous layer was concentrated to dryness and the resulting solid was recrystallized from acetone to provide 1-((methylamino)methyl)cyclopentanecarboxylic acid hydrobromide (127 mg, 48% yield) as a white crystalline solid.

1H-NMR (500 MHz, DMSO-de) 8 (ppm): 12.76-13.15 (s, 1 H), 8.12-8.39 (m, 2 H), 2.98-3, 11 (m, 2 H), 2.55 (s, 3 H), 1.86-2.01 (m, 2 H), 1.62 (m, 6 H).

Stage 4: Reference compound I-228

The title compound was prepared following general procedure B, except that 1-((methylamino)methyl)cyclopentanecarboxylic acid (as the HBr salt) was the amine reagent (1.3 equiv.), 4 equivalents of triethylamine, and the content was heated at 90 °C for 6 h as a solution in THF/water (10:1). The contents were extracted with ethyl acetate during processing. Purification was achieved by silica gel chromatography using 2 to 5% menthol in dichloromethane for 40 minutes. The desired compound was obtained as a white solid (13.4 mg, 45% yield).

1H NMR (400 MHz, CDCla) 8 (ppm): 8.44 (s, 1 H), 8.08 (d, 1 H), 7.29 (s, 1 H), 7.15 - 7.25 (m, 1 H), 6.95 - 7.08 (m, 3 H), 6.55-6.58 (m, 1 H), 5.95 (s, 2 H), 4.02 (s , 2 H), 3.35 (d, 3 H), 2 .18-2.29 (m, 2 H), 1.57- 1.79 (m, 6 H).

Reference compound I-238

The title compound was prepared in 4 steps:

Stage 1: 4-((4-methylphenylsulfonamido)methyl)tetrahydro-2H-pyran-4 carboxylic acid

A suspension of 4-(aminomethyl)tetrahydro-2H-pyran-4-carboxylic acid (500 mg, 1.0 equiv.), p-toluenesulfonyl chloride (719 mg, 1.2 equiv.) and aqueous solution of 1 M aqueous sodium hydroxide (9.4 mL, 3.0 equiv.) at 90 °C for 1 hour, after which the reaction mixture was cooled to 0 °C and acidified by the addition of an aqueous solution of 3 M hydrochloric acid. The resulting white precipitate was filtered, then washed successively with water and ethanol to provide 4-((4-methylphenylsulfonamido)methyl)tetrahydro-2H-pyran-4-carboxylic acid (840 mg, 85% yield). like a white solid. It was not necessary to purify.

1H-NMR (400 MHz, DMSO-de) 8 (ppm): 12.6 (br. s, 1 H), 7.68 (d, 2 H), 7.66 (t, 1 H), 7, 39 (d, 2 H), 3.63 - 3.72 (m, 2 H), 3.27 - 3.32 (m, 2 H), 2.81 (d, 2 H), 2.38 ( s, 3 H), 1.76 - 1.85 (m, 2 H), 1.33 - 1.46 (m, 2 H).

Stage 2: 4-((N,4-dimethylphenylsulfonamido)methyl)tetrahydro-2H-pyran-4-carboxylic acid

A suspension of 4-((4-methylphenylsulfonamido)methyl)tetrahydro-2H-pyran-4-carboxylic acid (840 mg, 1.0 equiv.) in a 1 M aqueous sodium hydroxide solution (10.7 ml, 4 .0 equiv.) and iodomethane (0.528 ml, 3.15 equiv.) was heated at 100 °C for two hours, after which the reaction mixture was diluted in 3 M aqueous hydrochloric acid solution, extracted with dichloromethane (3 x 30 ml), dried (sodium sulfate), filtered and concentrated to give 4-((N,4-dimethylphenylsulfonamido)methyl)tetrahydro-2H-pyran-4 carboxylic acid (197 mg, 22% yield). as a cream-colored solid. It was not necessary to purify.

1H NMR (500 MHz, CDCI3) 8 (ppm): 7.63 - 7.72 (d, 2 H), 7.31 - 7.39 (d, 2 H), 3.87 -3.97 (m , 2 H), 3.50 - 3.61 (m, 2 H), 3.25 (s, 2 H), 2.76 (s, 3 H), 2.45 (s, 3 H), 2 .13-2.23 (m, 2 H), 1.62 - 1.74 (m, 2 H).

Step 3: 4-((methylamino)methyl)tetrahydro-2H-pyran-4-carboxylic acid hydrobromide

A solution of 4-((N,4-dimethylphenylsulfonamido)methyl)tetrahydro-2H-pyran-4-carboxylic acid (197 mg, 1.0 equiv.) was heated in a 33% glacial acetic acid solution (1 ml , 31 equiv.) at 85 °C for 3 hours, after which an LCMS analysis indicated that the starting material had been consumed. After cooling the reaction mixture to room temperature, water was added, and the reaction mixture was washed with diethyl ether (3 x 30 ml). The aqueous layer was concentrated to dryness, and the resulting solid was recrystallized from acetone to obtain 4-((methylamino)methyl)tetrahydro-2H-pyran-4-carboxylic acid hydrobromide (54.8 mg, 36% yield) as a solid. white.

1H NMR (500 MHz, D2O) 8 (ppm): 3.71 - 3.89 (m, 2 H), 3.50-3.64 (m, 2 H), 3.17 (s, 2 H) , 2.66 (s, 3 H), 1.96-2.09 (m, 2 H), 1.48- 1.66 (m, 2 H).

Stage 4: Reference compound I-238

This compound was prepared following general procedure B, except that 4-((methylamino)methyl)tetrahydro-2 H-pyran-4-carboxylic acid (as the HBr salt) was the amine reagent (1.05 equiv. ), 4 equivalents of triethylamine were used, and the reaction was taken up in dioxane/water (3:1) at 90 °C for 18 hours. The contents were extracted with ethyl acetate during processing and purification was achieved by silica gel chromatography using 2 to 7% methanol in dichloromethane for 40 minutes providing the desired compound, reference compound I-238, as a solid. blank (31.0 mg, yield 43%) following the procedure described above.

1H NMR (400 MHz, CDCla) 8 (ppm): 8.47 (s, 1 H), 8.06 (d, 1 H), 7.31 (s, 1 H), 7.23 - 7.27 (m, 1 H), 7.22 (br. s, 1 H), 7.00 - 7.09 (m, 3 H), 6.59 (d, 1 H), 5.96 (s, 2 H), 3.83 - 3.95 (m, 4 H), 3.47 - 3.56 (m, 2 H), 3.40 (d, 3 H), 2.20 - 2.26 (m , 2 H), 1.51 -1.64 (m, 2 H).

Reference compound I-244

The title compound was prepared in 4 steps:

Step 1: 4,4,4-trifluoro-2-(4-methylphenylsulfonamido)butanoic acid

A suspension of 2-amino-4,4,4-trifluorobutanoic acid (300 mg, 1.0 equiv.), p-toluenesulfonyl chloride (437 mg, 1.2 equiv.) and aqueous sodium hydroxide solution was formed. 1 M (5.73 ml, 3.0 equiv.) in water (4 ml) at 90 °C for 1 hour, after cooling the reaction mixture to 0 °C and acidifying it by adding aqueous acid solution 3 M hydrochloric acid, extracted with dichloromethane (3 29% yield) as a white solid.

1H NMR (500 MHz, CDCla) 8 (ppm): 7.71 - 7.80 (d, 2 H), 7.28 (d, 2 H, isochronous with chloroform), 5.72 - 5.91 (br s, 1 H), 4.16 - 4.29 (m, 1 H), 2.64 - 2.76 (m, 1 H), 2.52 - 2.63 (m, 1 H), 2 .43 (s, 3 H).

Step 2: 2-(N,4-dimethylphenylsulfonylamido)-4,4,4-trifluorobutanoic acid

A mixture of 4,4,4-trifluoro-2-(4-methylphenylsulfonylamido)butanoic acid (175 mg, 1.0 equiv.) and iodomethane (146 pl, 3.15 equiv.) in aqueous sodium hydroxide solution 1 M (2.81 mL, 4.0 equiv.) was heated at 85 °C for 2.5 hours, after which LC/MS analysis indicated the presence of the desired product and the methyl ester of the desired product. The reaction mixture was acidified with 3M hydrochloric acid solution, extracted with dichloromethane (3 x 30 ml), dried (sodium sulfate), filtered and concentrated to a residue. This residue was reconstituted in tetrahydrofuran (2 ml) and then treated with a 1 M aqueous sodium hydroxide solution (0.5 ml). After 30 minutes of stirring at room temperature, the reaction mixture was acidified with 3 M hydrochloric acid solution, extracted with dichloromethane (3 x 30 ml), dried (sodium sulfate) and concentrated to yield acid 2 -(N,4-dimethylphenylsulfonylamido)-4,4,4-trifluorobutanoic acid (66 mg, 36% yield) as a gum with approximately 90% purity according to 1H NMR. It was used as such in the next step without further purification. 1H NMR (500 MHz, CDCh) 8 (ppm): 7.60 - 7.70 (d, 2 H), 7.19 (d, 2 H), 4.90 - 4.99 (m, 1 H) , 2.75 - 2.89 (m, 1 H), 2.66 -2.72 (s, 3 H), 2.30 - 2.44 (m, 1 H), 2.29 (s, 3 H).

Step 3: 4,4,4-trifluoro-2-(methylamino)butanoic acid hydrobromide

A solution of 2-(N,4-dimethylphenylsulfonylamido)-4,4,4-trifluorobutanoic acid (66 mg, 1.0 equiv.) in a solution of glacial acetic acid was heated to 33% hydrogen bromide (1, 0 ml, 91 equiv.) at 85 °C for 2 hours. There was still starting material left. It was allowed to stir at 60 °C for 72 hours, after which the deprotection was almost complete. The reaction mixture was diluted in water, washed with diethyl ether (3 x 30 ml), and the aqueous layer was concentrated to dryness. This crude material was used as is in the next step without purification.

Stage 4: Reference compound I-244

This compound was prepared following procedure B described above, except that 4,4,4-trifluoro-2-(methylamino)butanoic acid (as the HBr salt) was the amine reagent (1.2 equiv.), 4 equivalents of triethylamine and the reaction was taken up in dioxane/water (3:1) at 90 °C for 5 days. The crude material was purified by silica gel chromatography with a gradient of 2-10% methanol/dichloromethane over 40 minutes providing the desired compound, reference compound I-244 (24.7 mg, 32% yield) as a solid. tan in color.

1H NMR (500 MHz, CDCla) 8 (ppm): 8.77 (s, 1 H), 8.21 (d, 1 H), 7.41 (m, 1 H), 7.24 - 7.33 (m, 1 H), 6.07 - 7.13 (m, 1 H), 7.02 - 7.07 (m, 1 H), 6.90 (d, 1 H), 6.82-6 .88 (m, 1 H), 5.97 (s, 2 H), 3.38 - 3.46 (m, 2 H), 3.33 - 3.36 (m, 1 H), 3.03 - 3.19 (m, 3 H).

Reference compound I-94

The compound was prepared following general procedure B, except that methyl 1-aminocyclobutanecarboxylate was used as amine reagent, 5 equivalents of triethylamine were used, the reaction was heated to 90 °C as a solution in THF/water (10:1). for 14 h, followed by heating at 170 °C for 10 minutes in the microwave. The contents were then treated with water and solid 1N HCl and dried under vacuum. The crude material was purified by preparative reverse phase HPLC to provide the desired compound, reference compound I-94 (0.30 MG, 1.5% yield) as a white solid.

1H NMR (500 MHz, METHANOL-d4) 8 (ppm): 8.81 (d, 1 H), 8.23 (d, 1 H), 7.34 (s, 1 H), 7.25 - 7 .31 (m, 1 H), 7.01 - 7.13 (m, 2 H), 6 .86-6.94 (m, 2 H), 5.97 (s, 2 H), 2.89 (ddd, 2 H), 2.45-2.54(m, 2 H), 2 .07-2.14 (m, 1 H), 1.95-2.03 (m, 1 H).

Reference compound I-138

This compound was prepared as above with the exception that methyl 1-aminocyclopentanecarboxylate (as the HCl salt) was the amine reagent, the mixture was heated for 5 hours at 140 °C in DMA (volume: 142 |jl ) providing the ester. The reaction is then stirred at room temperature (23 °C) for 16 hours. Sodium hydroxide (14.2 mg) was added and reacted and heated at 40 °C for 1 h, then cooled, water was added, neutralized with 1N HCl and extracted with ethyl acetate (3 times ). The organic substances were combined and dried, purified by reverse phase preparative HPLC, obtaining the desired compound, the reference compound I-138 (0.5 mg, 1.5% yield).1

1H NMR (500 MHz, METHANOL-d4) 8 (ppm): 8.84 (s, 1 H), 8.29 (d, 1 H), 7.40 (s, 1 H), 7.28 - 7 .35 (m, 1 H), 7.04 - 7.16 (m, 2 H), 6.91 -7.00 (m, 2 H), 6.01 (s, 2 H), 2.50 -2.62 (m, 3 H), 2.17-2.26 (m, 2 H), 1.90 (br. s., 3 H).

Reference compound I-156

A mixture of intermediate 1 (30.8 mg), (1S,2R)-2-aminocyclopentanecarboxylic acid (31.9 mg, 3 equiv.) and triethylamine (115 jl, 10 equiv.) at 80 °C for 16 hours in a 10:1 THF/water mixture. The content was concentrated in vacuo and purified by reverse phase preparative HPLC to obtain the desired compound, compound I-156, as a white solid (6.2 mg, 16% yield). .

1H NMR (500 MHz, CDCI3) 5 ppm: 8.52 (s, 1 H), 8.46 (br. s., 1 H), 7.44 (br. s., 2 H), 7.22 - 7.27 (m, 1 H), 7.15 (t, 1 H), 7.00 - 7.09 (m, 2 H), 6.66 (s, 1 H), 5.94 (s , 2 H), 4.88 (br. s., 1 H), 3.13 - 3.21 (m, 1 H), 2.23 (d,1 H), 2.15 (br. s. , 2 H), 1.85-2.03 (m, 2 H), 1.76 (d, 1 H).

Reference compound I-154

The title compound was prepared following general procedure B, except that cis-2-aminocyclohexanecarboxylic acid was the amine reagent and the mixture was heated at 80 °C for 24 h as a solution in THF/water (10:1). . The content was concentrated in vacuo and purified by reverse phase HPLC to obtain the desired compound, reference compound I-154 (8.5 mg, 26% yield) as a white solid.

1H NMR (500 MHz, CDCla) 5 ppm: 8.62 (d, 1 H), 8.53 (d,1 H), 7.79 (br. s., 1 H), 7.45 (s, 1 H), 7.19 - 7.27 (m, 2 H), 7.00 - 7.10 (m, 2 H), 6.67 (s, 1 H), 5.94 (s, 2 H) ), 4.58 (br. s.,1 H), 2.94 (d, 1 H), 2.33 (d, 1 H), 1.87 (br. s., 2 H), 1, 81 (d, 1 H), 1.61 -1.74 (m, 2 H), 1.36 - 1.57 (m, 2 H).

Reference compound I-159

The title compound was prepared following general procedure B, except that 3-(4-hydroxyphenyl)-L-alanine was the amine reagent and the mixture was heated at 80 °C for 18 h as a solution in THF/water (10:1). The contents were concentrated in vacuo and purified by reverse phase HPLC to provide the desired compound, reference compound I-159 as a brown oil.

1H NMR (500 MHz, CD3OD) 5 ppm: 8.82 (d, 1 H), 8.22 (d, 1 H), 7.52 (s, 1 H), 7.25 - 7.33 (m , 1 H), 7.02 - 7.15 (m, 4 H), 6.97 (d, 1 H), 6.92 (t, 1 H), 6.64 (d, 2 H), 6 .00 (s, 2 H), 5.29 (dd, 1 H), 3.40 (dd, 1 H), 3.09 (dd, 1 H).

Reference compound I-165

The title compound was prepared following general procedure B, except that 3-(4-hydroxyphenyl)-D-alanine was the amine reagent and the mixture was heated at 80 °C for 90 h as a solution in THF/water (10:1). The contents were concentrated in vacuo and purified by reverse phase HPLC to provide the desired compound, reference compound I-165 (4.7 mg, 13% yield) as a brown oil.

1H NMR (500 MHz, CD3OD) 5 ppm: 8.82 (d, 1 H), 8.24 (d, 1 H), 7.53 (s, 1 H), 7.26 - 7.38 (m , 1 H), 7.13 (d, 2 H), 7.04 - 7.11 (m, 2 H), 6.98 (d, 1 H), 6.93 (t, 1 H), 6 .64 (d, 2 H), 6.01 (s, 2 H), 5.30 (dd, 1 H), 3.41 (dd, 1 H), 3.09 (dd, 1 H). Reference compound I-179

The title compound was prepared following general procedure B, except that (1S,3R)-3-aminocyclohexanecarboxylic acid was the amine reagent and the mixture was heated at 80 °C for 48 h as a solution in THF/water (10:1). The contents were concentrated in vacuo and purified by reverse phase HPLC to provide the desired compound, reference compound I-179 (1.7 mg, 5% yield).

1H NMR (400 MHz, CD3OD) 5 ppm: 8.83 (d, 1 H), 8.23 (d, 1 H), 7.67 (s, 1 H), 7.26 - 7.35 (m , 1 H), 7.12 (d, 1 H), 7.05 - 7.10 (m, 1 H), 7.01 (d, 1 H),) 6.94 - 7.00 (m, 1 H), 6.03 (s, 2 H), 2.96 - 3.06 (m, 1 H), 2.42 - 2.54 (m, 1 H), 2.21 (td,1 H) ), 1.97 - 2.15 (m, 4 H), 1.80- 1.96 (m, 1 H).

Reference compound I-188

The title compound was prepared following general procedure B, except that 4-fluoro-piperidinecarboxylic acid (as the HCl salt) was the amine reagent and the mixture was heated to 80 °C for 8 h as a solution in THF /water (10:1) followed by stirring at 23 °C for a further 8 h. The contents were concentrated in vacuo and purified by reverse phase HPLC to provide the desired compound, reference compound I-188 (7 mg, 18% yield) as a white solid.

1H NMR (500 MHz, CD3OD) 5 ppm: 8.81 (d, 1 H), 8.32 (d, 1 H), 7.63 (s, 1 H), 7.26 - 7.33 (m , 1 H), 7.03 - 7.14 (m, 2 H), 6.91 - 6.98 (m, 2 H), 6.01 (s, 2 H), 4.82 (br. s ., 1 H), 3.59-3.73 (m, 2 H), 2.26-2.41 (m, 2 H), 2.16-2.23 (m, 2 H), 0, 10 (m, 1 H).

Reference compound I-199

The title compound was prepared following general procedure B, except that (S)-2-amino-4-(methylmercapto)butyric acid was the amine reagent and the mixture was heated to 80 °C for 16 h as a solution. in THF/water (10:1). The content was concentrated in vacuo and purified by reverse phase HPLC to obtain the desired compound, reference compound I-199 (4 mg, 9% yield) as a white solid.1

1H NMR (500 MHz, CD3OD) 5 ppm: 8.82 (d, 1 H), 8.34 (d, 1 H), 7.58 (s, 1 H), 7.27 - 7.34 (m , 1 H), 7.04 -7.14 (m, 2 H), 6.93 - 7.00 (m, 2 H), 6.02 (s, 2 H), 5.24 (dd, 1 H), 2.59 - 2.79 (m, 2 H), 2.36 - 2.46 (m, 1 H), 2.22 - 2.31 (m, 1 H), 2.12 (s , 3H).

I-192 REFERENCE COMPOSITION

The title compound was prepared following general procedure B, except that 3-(methanesulfonyl)pyrrolidine was

amine reagent and the mixture was heated at 80 °C for 48 h as a solution in THF/water (10:1). The content

was acidified with 1N hydrochloric acid, concentrated in vacuo and purified by reverse phase HPLC providing

the desired compound, reference compound I-192 (6.3 mg, 18% yield) as a white solid.

1H NMR (500 MHz, CD3OD) 6 ppm: 8.81 (d, 1 H), 8.30 (d, 1 H), 7.61 (s, 1 H), 7.30 (ddd,1 H) , 7.03 - 7.14 (m, 2 H - 6.98 (m, 2 H), 6.01 (s, 2 H), 4.41 - 4.54 (m, 1 H), 4, 27 - 4.38 (m, 1 H), 4.06 - 4.27 (m, 3 H), 3.11 (s, 3 H), 2.52 - 2.68 (m, 2 H).

Reference compound I-220

The title compound was prepared following general procedure B, except that p-cyano-L-alanine was the reagent

of amine and the mixture was heated at 80 °C for 18 h as a solution in THF/water (10:1). The content was concentrated

in vacuo and purified by reverse phase HPLC to provide the desired compound, reference compound I-220 (2.5 mg, 8% yield) as a white solid.

1H NMR (500 MHz, CD3OD) 6 ppm: 8.79 (d, 1 H), 8.30 (d, 1 H), 7.52 (s, 7.25 - 7.31 (m, 1 H) , 7.02 - 7.13 (m, 2 H), 6.86 - 6.95 (m, 2 H), 5.99 (s, 2 H), 5.34 (dd, 1 H), 3 .15 - 3.25 (m, 2 H).

Reference compound I-198

The title compound was prepared following general procedure B, except that trans-2-aminocyclohexanecarboxylic acid was the amine reagent and the mixture was heated at 80 °C for 16 h as a solution.

in THF/water (10:1). The contents were acidified with 1 N hydrochloric acid solution, and the solids were filtered, resuspended in dichloromethane, and filtered to provide the desired compound, reference compound I-198 (14.5 mg, 31% yield). like a white solid.

1H NMR (500 MHz, METHANOL-d4) 6 ppm: 8.75 (d, 1 H), 8.00 (d, 1 H), 7.42 (s, 1 H), 7.23 - 7.29 (m, 1 H), 7.05 - 7.11

(m, 1 H), 7.02 (t, 1 H), 6.89 - 6.92 (m, 1 H), 6.81 (t, 1 H), 5.95 (s, 2 H) , 4.58 (td, 1 H), 2.56 (td, 1 H), 1.98 - 2.14 (m, 2

H), 1.78 - 1.90 (m, 2 H), 1.67 (qd, 1 H), 1.48 - 1.61 (m, 1 H), 1.28 - 1.47 (m , 2H).

Reference compound I-208

The title compound was prepared following general procedure B, except that octahydrocyclopenta[c]pyrrole-3a-carboxylic acid (4 equiv.) was the amine reagent and the mixture was heated to 80 °C for 5 h as a solution in THF/water (10:1). The contents were dried by blowing with nitrogen, and the crude mixture was resuspended in methanol and filtered to provide the desired compound, reference compound I-208 (37 mg, 93% yield).

1H NMR (500 MHz, METHANOL-d4) 6 ppm: 8.74 (d, 1 H), 8.06 - 8.13 (m, 1 H), 7.39 - 7.45 (m, 1 H) , 7.26 (m, 1H), 7.09

(m, 1 H), 7.02 (d, 1 H), 6.91 (d, 1 H), 6.82 (m, 1 H), 5.96 (s., 2 H), 4, 40 (d, 1 H), 4.06 (m, 1 H), 3.79 (d, 2 H), 3.06 (br.

s., 1 H), 2.31 (m, 1 H), 2.11 (m, 1 H), 1.90 (m, 2 H), 1.64 (m, 1 H),1.30 (m, 1 H).

Reference compound I-233

The title compound was prepared following general procedure B, except that L-cyclohexylglycine methyl ester (as the HCl salt) was the amine reagent, and the contents were heated to 90 °C as a solution in THF/water ( 10:1) for 16 h. The contents were cooled, treated with solid sodium hydroxide and stirred at 23 °C for

2 hours The organic solvent was removed from the reaction mixture upon completion and the precipitate was filtered to provide the desired compound, reference compound I-233 as a white solid (26.0 mg, 0.047 mmol, yield 70.7

%).

1H NMR (500 MHz, METHANOL-d4) 6 ppm: 9.08 (d,1 H), 8.12 (d, 1 H), 7.42 (s, 1 H), 7.29-7.35 (m, 1 H), 7.25 (d, 1 H),

7.18-7.24 (m, 1 H), 7.09 (t, 1 H), 6.93 (t, 1 H), 6.77 (d, 1 H), 5.82-5, 92 (dd, 2 H), 4.17 (br. s., 1 H), 3.30 (s., 1 H), 1.79 -1.91 (m, 2 H), 1.50 - 1.69 (m, 3 H), 0.89 - 1.24 (m, 5 H).

Reference compound I-243

A mixture of intermediate 1 (25 mg, (S)-methyl-2-amino-2-cyclohexyl acetate hydrochloride (41.7 mg, 3 equiv.) and triethylamine (93 pl, 10 equiv.), was heated to 90° C for 16 cycles in a THF/water mixture.

cooled, NaOH (5.35 mg, 2 equiv) was added, and the mixture was stirred at room temperature for 2 h. The organic solvent was removed and the resulting precipitate was filtered to provide acid (R)-2-cyclohexyl-2-((5-fluoro-2-(1-(2-fluorobenzyl)-5-(isoxazol-3-yl)- 1 H-pyrazol-3-yl)pyrimidin-4-yl)amino)acetic acid as a white solid (26.0 mg, 0.047 mmol, 70.7% yield).12

1H NMR (500 MHz, METHANOL-d4) 6 ppm: 8.75 (d, 1 H), 8.11 (d, 1 H), 7.41 (s, 1 H), 7.23 - 7.29 (m, 1 H), 7.00 - 7.11

(m, 1 H), 7.02 (t, 1 H), 6.88 (d, 1 H), 6.83 (t, 1 H), 5.95 (s, 2 H), 4.73 (d,1 H), 1.97-2.04 (m, 1 H), 1.88 (t, 2 H), 1.80 (d,

2 H), 1.70 (d, 1 H), 1.17 -1.39 (m, 5 H). The title compound was also prepared following the general procedure

B, except that methyl D-cycIohexylglycine methyl ester (as the HCl salt) was the amine reagent (1 equiv.), and the contents were heated to 90 °C for 16 h as a solution in THF/water (10 :1). The contents were cooled, treated with solid sodium hydroxide and stirred at 23 °C for 18 h. The content was concentrated in vacuo and purified by reverse phase HPLC to provide the desired compound, reference compound I-243 (1 mg, 3% yield) as a white solid.

Reference compound I-242

The title compound was prepared in 4 steps:

Stage 1: trans-2-(4-methylphenylsulfonamido)cyclohexanecarboxylic acid

A suspension of trans-2-aminocyclohexanecarboxylic acid (318 mg, 1.0 equiv.), ptoluenesulfonyl chloride (508 mg, 1.2 equiv.) and 1 M aqueous sodium hydroxide solution (6.7 ml, 3.0 equiv) in water (5 ml) at 90 °C for 1 hour. The reaction mixture was cooled to 0 °C, and acidified by adding a 3 M aqueous solution of hydrochloric acid. The resulting white precipitate was filtered and washed successively with water and then ethanol to provide trans-2-(4 Racemic -methylphenylsulfonamido)cyclohexanecarboxylic acid as a white solid (179.6 mg, 27% yield).

1H NMR (400 MHz, CDCh) 8 (ppm): 7.73 (d, 2 H), 7.27 (d, 2 H), 4.98 - 5.16 (m, 1 H), 3.24 - 3.46 (br. s, 1 H), 2.39 (s, 3 H), 2.23 - 2.34 (m, 1 H), 1.87 - 2.03 (m, 2 H) , 1.58 - 1.78 (m, 2 H), 1.42 - 1.58 (m, 1 H), 1.08 - 1.35 (m, 3 H).

Stage 2: Synthesis of frans-2-(N,4-dimethylphenylsulfonamido)cyclohexanecarboxylic acid

A solution of trans-2-(4-methylphenylsulfonamido)cyclohexanecarboxylic acid (187 mg, 0.629 mmol), iodomethane (0.124 mL, 3.0 equiv), and 1 M aqueous sodium hydroxide solution (2.52 mL, 4.0 equiv) in water (5 ml) was heated at 75 °C for 1.5 hours, after which the reaction mixture was cooled to room temperature, washed with dichloromethane (2 x 30 ml), acidified by the addition of 3 M aqueous hydrochloric acid solution, extracted with dichloromethane (3 x 30 ml), dried (sodium sulfate), filtered, and concentrated to give the crude N-methylamino acid product. Purification was achieved using silica gel chromatography with 2 to 5% methanol in dichloromethane as eluent over the course of 40 minutes. This gave trans-2-(A/,4-dimethylphenylsulfonamido)cyclohexanecarboxylic acid as a white foam (130 mg, 66% yield).

1H NMR (500 MHz, CDCla) 8 ppm: 7.69 - 7.76 (d, 2 H), 7.28 (d, 2 H), 4.03 - 4.16 (m, 1 H), 2 .78 (s, 3 H), 2.49 - 2.61 (m, 1 H), 2.43 (s, 3 H), 2.02-2.13 (m, 1 H), 1.73 - 1.84 (m, 2 H), 1.63- 1.73 (m, 1 H), 1.55- 1.63 (m, 1 H), 1.35- 1.45 (m, 2 H), 1.10-1.22 (m, 1 H).

Step 3: trans-2-(methylamino)cyclohexanecarboxylic acid hydrobromide

To a vial containing trans-2-(N,4-dimethylphenylsulfonylamido)cyclohexanecarboxylic acid (130 mg, 1.0 equiv) was added a 33% glacial acetic acid solution (1.2 mL, 53 equiv). The suspension was heated at 85 °C for 2.5 hours, after which it was diluted with water and washed with diethyl ether (2 x 30 ml), then concentrated to a golden foamy residue. Recrystallization of this material from acetone provided trans-2-(methylamino)hexanecarboxylic acid hydrobromide as a cream solid (54.4 mg, 55% yield).

1H NMR (500 MHz, D2O) 8 (ppm): 3.24 - 3.37 (m, 1 H), 2.65 (s, 3 H), 2.47 - 2.60 (m, 1 H) , 2.06 - 2.20 (m, 2 H), 1.76 -1.84 (m, 1 H), 1.15-1.51 (m, 5 H).

Stage 4: Reference compound I-242

The title compound was prepared following general procedure B, except that trans-2-(methylamino)cyclohexanecarboxylic acid (as the HBr salt) was the amine reagent, and the contents were heated at 85 °C for 18 h as a solution in THF/water (10:1). The contents were cooled, concentrated in vacuo and purified by reverse phase HPLC to provide the desired compound, reference compound I-242 (1 mg, 3% yield) as a white solid.

1H NMR (500 MHz, M ETAN O L^) 8 ppm: 8.75 (d, 1 H), 8.09 (d, 1 H), 7.38 (o, 1 H), 7.23 - 7, 30 (m, 1H), 7.06 - 7.11

(m, 1 H), 7.00 - 7.05 (m, 1 H), 6.89 (d, 1 H), 6.83 (t, 1 H), 5.94 (o, 2 H) , 3.16 - 3.24 (m, 3 H), 2.79 (br. s., 1 H), 2.08 (d,

1 H), 1.86 - 1.97 (m, 2 H), 1.81 (d, 2 H), 1.45 - 1.70 (m, 2 H), 1.34 (dt, 1 H )

Reference compound I-31

The title compound was prepared following general procedure B, except that 2-amino-1-morpholinoethanone (5 equiv.) was the amine reagent, and 3 equivalents of triethylamine were used, and the contents were heated to 80 °C for 1 h as a solution in THF. The solvent was removed in vacuo and the contents taken up in ethyl acetate.

The organic layer was washed with 1 N hydrochloric acid solution, water and brine, dried with sodium sulfate, filtered and concentrated in vacuo. The crude material was purified by silica gel chromatography using a gradient of

0 to 100% ethyl acetate/hexane providing the desired compound, reference compound I-31 (4.7 mg, 23% yield).

1H NMR (500 MHz, CDCh) 88.44-8.52 (m, 1 H), 8.14-8.25 (m, 1 H), 7.19-7.27 (m, 1 H), 6.97-7.12 (m, 2 H), 6.83-6.91

(m, 1 H), 6.61-6.66 (m, 1 H), 6.00 (s, 2 H), 4.39-4.47 (m, 2 H), 3.71-3 .82 (m, 7 H), 3.56-3.63 (m, 2 H).

Reference compound I-33

The title compound was prepared following general procedure B, except that 3-methylmorpholine was the amine reagent, 5 equivalents of triethylamine were used, and the contents were heated to 60 °C for 18 h as a solution in THF, and then at 80 °C for 18 h. The solvent was removed in vacuo and the contents taken up in ethyl acetate. The organic layer was washed with 1N hydrochloric acid, water and brine, dried with sodium sulfate, filtered and concentrated in vacuo. The crude material was purified by silica gel chromatography with a gradient of 0-5% methanol/dichloromethane providing the reference compound I-33 (8 mg, 41% yield).

1H NMR (500 MHz, CDCla) 88.48 (m, 1 H), 8.22 (m, 1 H), 7.31 (s, 1 H), 7.1822 (m, 1 H), 6, 99 (s, 2 H), 6.87 (m, 1 H) 6.61 (d, 1 H), 5.98 (s, 2 H), 4.69 (m, H), 4.37 (m , 1 H), 4.05 (m, 1 H), 3.83 (m, 2 H), 3.69 (m, 1 H), 3.4752 (m, 1 H), 1.45 (d , 3H).

Reference compound I-34

The title compound was prepared following general procedure B, except that methyl pyrrolidine-2-carboxylate was the amine reagent, 2 equivalents of triethylamine were used, and the contents were heated at 60 °C for 18 h as a solution. in THF. The solvent was removed in vacuo and the contents taken up in ethyl acetate. The organic layer was washed with 1 N hydrochloric acid solution, water and brine, dried with sodium sulfate, filtered and concentrated in vacuo. The crude material was purified by silica gel chromatography with a gradient of 0-5% methanol/dichloromethane providing the desired compound, reference compound I-34 (10.6 mg, yield

57%).

1H NMR (500 MHz, CDCh) 88.47 (m, 1 H), 8.20 (m, 1 H), 8.17.29 (s, 1 H), 7.21 (m, 1 H), 7.04 (m, 1H), 6.98 (m, 1H),

6.87 (m, 1 H), 6.59 (dm, 1 H), 5.98 (m, 2 H), 4.76 (m, 1 H), 4.05 (m, 1 H), 3.94 (m, 1 H), 3.73 (s, 3 H), 2.35 (m, 1 H),

2.17 (m, 3 H).

Reference compound I-35

The title compound was prepared following general procedure B, except that tert-butylpyrrolidin-3-ylcarbamate was the amine reagent (5 equiv.), 3 equivalents of triethylamine were used, and the contents were heated to 80 °C for

1 h as a solution in<t>H<f>. The solvent was removed in vacuo and the contents were purified by silica gel chromatography using a gradient of 0-100% ethyl acetate/hexanes providing the desired compound, reference compound I-35 (30 mg, 68% yield). .

1H NMR (500 MHz, CDCh) 88.44-8.48 (m, 1 H), 8.14-8.19 (m, 1 H), 7.32 (s, 1 H), 7.17- 7.24 (m, 1 H), 6.95-7.08 (m, 2

H), 6.82-6.89 (m, 1 H), 6.57-6.63 (m, 1 H), 5.99 (s, 2 H), 4.72-4.79 (m , 1 H), 4.32-4.43 (m, 1 H), 4.00-4.07 (m, 1 H),

3.86-3.95 (m, 2 H), 3.68-3.75 (m, 1 H), 2.23-2.33 (m, 1 H), 1.96-2.05 ( m, 1 H), 1.48 (s, 9 H).

REFERENCE COMPOSITION I-41

The title compound was prepared following general procedure B, except that methyl pyrrolidine-2-carboxylate was the amine reagent, 2 equivalents of triethylamine were used, and the contents were heated at 60 °C for 18 h as a solution. in THF. The solvent was removed in vacuo and the contents taken up in ethyl acetate. The organic layer was washed with 1 N hydrochloric acid solution, water and brine, dried with sodium sulfate, filtered and concentrated in vacuo. The crude material was purified by silica gel chromatography with a gradient of 0-5% methanol/dichloromethane providing the reference compound I-41 (4 mg, 22% yield).

1H NMR (500 MHz, CDCh) 88.44-8.49 (m, 1 H), 8.19-8.26 (m, 1 H), 7.34-7.39 (m, 1 H), 7.18-7.25 (m, 1H), 6.92-7.10

(m, 3 H), 6.74-6.80 (m, 1 H), 5.95-6.00 (m, 2 H), 4.45-4.51 (m, 2 H), 2 .42-2.51 (m, 3 H), 2.16-2.23 (m, 4 H).

Reference compound I-46

The title compound was prepared by treating a solution of reference compound I-35 in dichloromethane with an equal volume of trifluroacetic acid. After stirring at 23 °C for 1 h, the solvent was removed under a stream of nitrogen, and the contents were dried in vacuo for 18 h to provide the desired compound, reference compound I-46 (29 mg) as a solid. .

1H NMR (500 MHz, CD3OD) 88.83-8.87 (m, 1 H), 8.37-8.42 (m, 1 H), 7.59-7.63 (m, 1 H), 7.29-7.37 (m, 1 H), 7.05-7.16

(m, 2 H), 6.94-7.02 (m, 2 H), 6.04 (s, 2 H), 4.13-4.33 (m, 5 H), 2.53-2 .64 (m, 1 H), 2.27-2.39 (m, 1 H).

Reference compound I-48

The title compound was prepared following general procedure B, except that methyl piperidine-2-carboxylate was the amine reagent, 2 equivalents of triethylamine were used, and the contents were heated at 60 °C for 18 h as a solution. in THF. The solvent was removed in vacuo and the contents taken up in ethyl acetate. The organic layer was washed with 1 N hydrochloric acid solution, water and brine, dried with sodium sulfate, filtered and concentrated in vacuo. The crude material was purified by reverse phase HPLC providing the desired compound, reference compound I-48 (3.7 mg, 18% yield).

1H NMR (400 MHz, CDCla) 88.46-8.50 (m, 1 H), 8.37-8.44 (m, 1 H), 7.32-7.37 (m, 1 H), 7.17-7.23 (m, 1 H), 6.97-7.09

(m, 3 H), 6.59-6.62 (m, 1 H), 5.91 (s, 2 H), 5.46-5.57 (m, 1 H), 4.54-4 .67 (m, 1 H), 3.75 (s, 3 H), 3.38-3.47 (m, 1 H), 2.34-2.45 (m, 1 H), 1.78 -1.89 (m, 3 H), 1.61-1.72 (m, 1 H), 1.45-1.55 (m, 1 H).

Reference compound I-53

The title compound was prepared following general procedure B, except that acetidine-3-carboxylic acid was the amine reagent (5 equiv.), 3 equivalents of triethylamine were used, and the contents were heated at 75 °C for 18 h as a solution in THF. The solvent was removed under a stream of nitrogen. The product was isolated by

Reversed phase HPLC providing the desired compound, reference compound I-53 (15.4 mg, yield

88%).

1H NMR (500 MHz, METHANOL-d4) 88.81-8.85 (m, 1 H), 8.22-8.27 (m, 1 H), 7.55-7.59 (m, 1 H ), 7.29-7.36 (m, 1 H), 7.05-7.16 (m, 2 H), 6.93-6.99 (m, 2 H), 5.99-6, 05 (m, 2 H), 4.65-4.84 (m, 4 H), 3.75-3.84 (m, 1 H).

Reference compound I-54

The title compound was prepared following general procedure B, except that 3-methylpiperazin-2-one was the amine reagent (5 equiv.), 3 equivalents of triethylamine were used, and the contents were heated to 75 °C for 18 h as a solution in THF. The solvent was removed under a stream of nitrogen. The product was isolated by reverse phase HPLC providing the desired compound, reference compound I-54 (1.4 mg, 8% yield).

1H NMR (500 MHz, CDCh) 88.53-8.55 (m, 1 H), 8.49-8.53 (m, 1 H), 7.43-7.48 (m, 1 H), 7.31-7.37 (m, 1 H), 7.24-7.28

(m, 1 H), 7.09-7.14 (m, 1 H), 7.02-7.08 (m, 2 H), 6.67-6.70 (m, 1 H), 5 .97 (s, 2 H), 5.34-5.47 (m, 1 H), 4.89-4.95 (m, 1

H), 3.62-3.78 (m, 2 H), 3.50-3.60 (m, 1 H), 1.70 (d, 3 H).

Reference compound I-55

The title compound was prepared following general procedure B, except that acetidine-2-carboxylic acid (5 equiv.) was the amine reagent, and 5 equivalents of triethylamine were used, and the contents were heated to 75 °C for 18 h as a solution in THF. The solvent was removed in vacuo and the contents were purified by reverse phase HPLC to provide the desired compound, reference compound I-55 (1.3 mg, 2% yield).

<1>H R M N (500 M Hz, C D<3>O D ) 8 8 .82 (s, 1 H), 8 .23 -8 .29 (m, 1 H), 7 .40 -7 .52 (m, 1 H), 7 .28 -7 .35 (m, 1 H), 7 .04 -7 .16 (m,

2 H), 6 .93 (br. s., 2 H), 5 .98 -6 .03 (m , 2 H), 5 .23 -5 .36 (m, 1 H), 4 .42 -4 .67 (m, 2 H), 2 .92 -3 .07 (m, 1 H), 2 .50 -2 .62 (m, 1

H).

I-56 REFERENCE COMPOSITION

The title compound was prepared following general procedure B, except that 3-fluoropiperidine (5 equiv.) was the amine reagent, 5 equiv. of triethylamine was used, and the contents were heated to 75 °C for 18 h as a solution. in THF. The solvent was removed in vacuo and the contents were purified by reverse phase HPLC to provide the desired compound, reference compound I-56 (1.3 mg, 2% yield).

1H NMR (500 MHz, CHLOROFORM-d) 88.52-8.56 (m, 1 H), 8.45-8.50 (m, 1 H), 7.49-7.54 (m, 1 H) ), 7.24-7.28 (m, 1

H), 7.13-7.20 (m, 1 H), 7.01-7.11 (m, 2 H), 6.68 (s, 1 H), 5.95 (s, 2 H) , 4.85-5.03 (m, 1 H), 4.56-4.81 (m, 2 H), 3.71-3.89

(m, 1 H), 3.47-3.60 (m, 1 H), 1.75-2.26 (m, 4 H).

Reference compound I-57

The title compound was prepared following general procedure B, except that 3,3-difluoropiperidine (5 equiv.) was the amine reagent, 5 equiv. of triethylamine was used, and the contents were heated at 75°C for 18 h. as a solution in THF. The solvent was removed in vacuo and the contents were purified by reverse phase HPLC to provide the desired compound, reference compound I-57 (4.5 mg, 5% yield).

1H NMR (500 MHz, CDCh) 88.52-8.55 (m, 1 H), 8.47-8.52 (m, 1 H), 7.40-7.45 (m, 1 H), 7.24-7.28 (m, 1 H), 7.11-7.17

(m, 1 H), 7.02-7.10 (m, 2 H), 6.65-6.68 (m, 1 H), 5.93-5.98 (m, 2 H), 4 .20-4.30 (m, 2 H), 4.00-4.08 (m, 2 H), 2.16-2.27

(m, 2 H), 1.96-2.05 (m, 2 H).

Reference compound I-58

A solution of reference compound I-48 was dissolved in THF, and an aqueous solution of lithium hydroxide (3 equiv.) was added. The solution was stirred at 25 °C for 18 h. The content was concentrated and the remaining aqueous layer was acidified with a 1 N hydrochloric acid solution, giving rise to a white precipitate. The aqueous layer was extracted with ethyl acetate and the combined organic layers were washed with water and brine. The contents were dried with sodium sulfate, filtered and concentrated to provide the desired compound, reference compound I-58 (29 mg, 100% yield).

1H NMR (400 MHz, CD3OD) 88.75 (m, 1 H), 8.20 (m, 1 H), 7.42 (m, 1 H), 7.26 (m, 1 H), 6. 98-7.11 (m, 2H), 6.84 (m,

2 H), 5.95 (s, 2 H), 5.47 (m, 1 H), 4.52 (m, 1 H), 3.44 (m, 1 H), 2.31-2, 40 (m, 1 H), 1.93 (m, 1 H), 1.81 (m, 2 H), 1.68

(m, 1 H), 1.54 (m, 1 H).

Reference compound I-59

The title compound was prepared following general procedure B, except that piperazin-2-one was the amine reagent (5 equiv.), 5 equivalents of triethylamine were used, and the contents were heated at 75 °C for 18 h. as a solution in THF. The solvent was removed under a stream of nitrogen and the content was purified by reverse phase HPLC to provide the desired compound, reference compound I-59 (1.6 mg, 2% yield).

1H NMR (500 MHz, CDCla ) 88.51-8.54 (m, 1 H), 8.45-8.49 (m, 1 H), 7.69-7.73 (m, 1 H), 7.23-7.27 (m, 1 H), 7.03-7.09

(m, 3 H), 6.69-6.73 (m, 2 H), 6.00-6.03 (m, 2 H), 4.70-4.73 (m, 2 H), 4 .26-4.32 (m, 2 H), 3.64-3.69 (m, 2 H).

Reference compound I-60

The title compound was prepared following general procedure B, except that triethylamine was the amine reagent and the content was heated to 60 °C for 18 h as a solution in THF. The solvent was removed in vacuo and the contents taken up in ethyl acetate. The organic layer was washed with 1 N hydrochloric acid solution, water and brine, dried with sodium sulfate, filtered and concentrated in vacuo. The crude material was purified by HPLC

reverse phase providing the desired compound, the reference compound I-60 (1.9 mg, 11% yield).

1H NMR (400 MHz, CD3OD) 88.77-8.81 (m, 1 H), 8.17-8.22 (m, 1 H), 7.48-7.52 (m, 1 H), 7.23-7.32 (m, 1 H), 7.01-7.13

(m, 2 H), 6.89-6.98 (m, 2 H), 5.96-6.01 (m, 2 H), 3.81-3.90 (m, 4 H), 1 .34 (s, 6 H).

Reference compound I-66

The title compound was prepared following general procedure B, except that piperidine-3-carboxamide was the amine reagent (5 equiv.), 8 equivalents of triethylamine were used, and the contents were heated at 75 °C for 18 h. as a solution in THF. The solvent was removed and the crude material was purified by reverse phase HPLC to provide the desired compound, reference compound I-66 (7 mg, 36% yield).

1H NMR (500 MHz, CDCh) 8 8.51-8.55 (m, 1 H), 8.25-8.31 (m, 1 H), 7.47-7.52 (m, 1 H) , 7.22-7.27 (m, 1 H), 7.09-7.16

(m, 1 H), 7.00-7.09 (m, 2 H), 6.83-6.90 (m, 1 H), 6.67-6.72 (m, 1 H), 6 .17-6.22 (m, 1 H), 5.90-5.95 (m, 2 H), 4, (m, 1 H), 4.30-4.43 (m, 1 H), 3.81-3.90 (m, 1 H), 3.60-3.69 (m, 1 H), 2.69-2.81 (m, 1 H), 2.05-2.13 ( m, 2H), 1.92-2.00

(m, 1 H), 1.69-1.81 (m, 1 H).

I-75 REFERENCE COMPOSITION

The title compound was prepared following general procedure B, except that methyl acepan-2-carboxylate was the amine reagent (1.5 equiv.), potassium carbonate (4 equiv.) was used instead of triethylamine, and the content was heated at 150 °C for 10 min in the microwave as a solution in NMP. The resulting mixture was filtered to remove solid potassium carbonate and concentrated in vacuo. The crude material was purified by reverse phase HPLC using a gradient of 20-70% acetonitrile/water (with 0.1% TFA) providing the desired compound, reference compound I-75 (1 mg, 3% yield). .

1H NMR (500 MHz, CDCla) 88.83 (m, 1 H), 8.33 (m, 1 H), 7.48 (m, 1 H), 7.31 (m, 1 H), 7. 10 (m, 2 H), 6.91 (m, 2 H), 6.01 (s, 2 H), 5.04 (m, 1 H), 4.18 (m, 1 H), 3, 73 (m, 1 H), 2.58 (m, 1 H), 2.04 (m, 3 H), 1.92 (m, 1 H), 1.79 (m, 1 H), 1, 53 (m, 2 H).

Reference compound I-82

The title compound was prepared following general procedure B, except that (1R,3S,5S)-8-azabicyclo[3.2.1]octan-3-yl-methanol (as the HCl salt) was the amine reagent. (3.5 equiv.), 5 equivalents of triethylamine were used, and the content was heated at 120 °C for 30 min in the microwave as a solution in NMP. The resulting mixture was purified by reverse phase HPLC to provide the desired compound, reference compound I-82 (10.8 mg, 42% yield).

1H NMR (500 MHz, METHANOL-d4) 88.82-8.86 (m, 1 H), 8.25-8.29 (m, 1 H), 7.62-7.66 (m, 1 H) ), 7.30-7.36 (m, 1 H), 7.05-7.15 (m, 2 H), 6.92-7.02 (m, 2 H), 6.00-6, 06 (m, 2 H), 3.66-3.72 (m, 2 H), 2.10-2.40 (m, 4 H), 1.78-2.07 (m, 5 H).

Reference compound I-83

The title compound was prepared following general procedure B, except that morpholine-2 carboxylic acid (as the HCl salt) was the amine reagent (2 equiv.), Hunig's base (3 equiv.) was used instead. of triethylamine, and the content was heated to 120 °C for 30 min in the microwave as a solution in NMP. The resulting mixture was purified by reverse phase HPLC using a gradient of 0-95% acetonitrile/water (with 0.1% TFA) to provide the desired compound, reference compound I-83 (10.8 mg, yield 42 %) like transparent glass.

1H NMR (500 MHz, METHANOL-d4) 88.81-8.85 (m, 1 H), 8.35-8.44 (m, 1 H), 7.64 (s, 1 H), 7. 26-7.35 (m, 1 H), 7.04 7.14 (m, 2 H), 6.97 (d, 2 H), 6.02 (s, 2 H), 4.73 (m , 1 H), 4.46 (m, 2 H), 4.14-4.20 (m, 1 H), 3.97 (m, 1 H), 3.89 (d, 2 H).

Reference compound I-87

The title compound was prepared following general procedure B, except that (R)-piperidine-2-carboxylic acid (4 equiv.) was the amine reagent, 5 equivalents of triethylamine were used, and the contents were heated to 90 °C for 18 h as a solution in THF/water (9:1). The solvent was removed under a stream of nitrogen, and the crude material was purified by reverse phase HPLC using a gradient of 20-51% acetonitrile/water (in 0.1% TFA) providing the desired compound, the reference compound. I-87 (12 mg, yield 48%).

1H NMR (500 MHz, METHANOL-d4) 88.79-8.83 (m, 1 H), 8.34-8.39 (m, 1 H), 7.60 (s, 1 H), 7. 27-7.35 (m, 1 H), 7.03 7.15 (m, 2 H), 6.90-6.98 (m, 2 H), 6.02 (s, 2 H), 4 .61-4.83 (m, 1 H), 3.43-3.58 (m, 1 H), 2.43-2.51 (m, 1 H), 1.69-2.02 (m , 5 H), 1.55-1.69 (m, 1 H).

Reference compound I-84

The title compound was prepared following general procedure B, except that (S)-piperidine-2-carboxylic acid (4 equiv.) was the amine reagent, 5 equivalents of triethylamine were used, and the contents were heated to 90 °C for 18 h as a solution in THF/water (9:1). The solvent was removed under a stream of nitrogen, and the crude material was purified by reverse phase HPLC using a gradient of 20-51% acetonitrile/water (in 0.1% TFA) providing the desired compound, reference compound I. -84 (9.6 mg, yield 39%).

1H NMR (500 MHz, METHANOL-d4) 88.79-8.83 (m, 1 H), 8.31-8.36 (m, 1 H), 7.54-7.58 (m, 1 H) ), 7.27-7.34 (m, 1 H), 7.04-7.15 (m, 2 H), 6.89-6.97 (m, 2 H), 6.01 (s, 2 H), 5.65 (br. s., 1 H), 4.58-4.80 (m, 1 H), 3.42-3.57 (m, 1 H), 2.41 2, 50 (m, 1 H), 1.67-2.02 (m, 4 H), 1.55-1.66 (m, 1 H).

Reference compound I-95

The title compound was prepared following general procedure B, except that (R)-morpholine-3-carboxylic acid (4 equiv.) was the amine reagent, Hunig's base (5 equiv.) was used instead of triethylamine. , and the content was heated to 90 °C for 18 h as a solution in THF/water (9:1). The solvent was removed under a stream of nitrogen, and the crude material was purified by reverse phase HPLC using a gradient of 20-51% acetonitrile/water (in 0.1% TFA) providing the desired compound, reference compound I. -95 (19 mg, 76% yield).

<1>H R M N (500 M Hz, C D<3>O D ) 8 8.81 (d, 1 H), 8 .39 (d, 1 H), 7 .57 (s, 1 H), 7 .26 - 7 .34 (m, 1 H), 7 .02 - 7 .16 (m, 2 H), 6 .93 (d, 2 H), 6 .00 (s, 2 H), 5 .26 - 5 , 59 (m, 1 H), 4 .55 (d, 2 H), 4 .04 (s, 1 H), 3 .93 (dd, 1 H), 3 .62 - 3 .80 (m, 2 H ).

I-96 REFERENCE COMPOSITION

The title compound was prepared following general procedure B, except that (S)-morpholine-3-carboxylic acid (4 equiv.) was the amine reagent, Hunig's base (5 equiv.) was used instead of triethylamine , and the content was heated to 90 °C for 18 h as a solution in THF/water (9:1). The solvent was removed under a stream of nitrogen, and the crude material was purified by reverse phase HPLC using a gradient of 20-51% acetonitrile/water (in 0.1% TFA) providing the desired compound, reference compound I. -96 (8 mg, yield 31%).

1H NMR (500 MHz, METHANOL-d4) 88.81 (s, 1 H), 8.34-8.42 (m, 1 H), 7.51-7.59 (m, 1 H), 7, 27-7.35 (m, 1 H), 7.02 7.15 (m, 2 H), 6.94 (s, 2 H), 6.01 (s, 2 H), 5.37-5 .54 (m, 1 H), 4.56 (d, 2 H), 4.01-4.09 (m, 1 H), 3.89-3.96 (m, 1 H), 3.61 -3.81 (s, 2 H).

Reference compound I-97

The title compound was prepared following general procedure B, except that 1,2,3,4-tetrahydroisoquinoline-1-carboxylic acid (4 equiv.) was the amine reagent, Hunig's base (5 equiv.) was used. instead of triethylamine, and the content was heated to 90 °C for 18 h as a solution in THF/water (9:1). The solvent was removed under a stream of nitrogen, and the contents were taken up in ethyl acetate. The organic layer was washed with 1 N hydrochloric acid solution, water and brine, dried with sodium sulfate, filtered and concentrated in vacuo. The crude material was purified by reverse phase HPLC to provide the desired compound, reference compound I-97 (3.4 mg, 12% yield) as a solid.

1H NMR (500 MHz, METHANOL-d4)88.84-8.89 (m, 1 H), 8.42-8.49 (m, 1 H), 7.62-7.69 (m, 2 H ), 7.30-7.40 (m, 4 H), 7.06-7.16 (m, 2 H), 6.97 (m, 2 H), 6.12-6.18 (m, 1H), 6.05 (s, 2H), 4.48-4.58 (m, 1H), 4.14-4.23 (m, 2H), 3.05-3.15 (m , 1 HOUR).

Reference compound I-98

The title compound was prepared following general procedure B, except that 3-methyl-5-(piperidin-2yl)-1,2,4-oxadiazole (4 equiv.) was the amine reagent, base of Hunig (5 equiv.) instead of triethylamine, and the content was heated to 90 °C for 18 h as a solution in THF/water (9:1). The solvent was removed under a stream of nitrogen, and the contents were taken up in ethyl acetate. The organic layer was washed with 1 N hydrochloric acid solution, water and brine, dried with sodium sulfate, filtered and concentrated in vacuo. The crude material was purified by reverse phase HPLC to provide the desired compound, compound I-98 (5.3 mg, 20% yield) as a solid.

1H NMR (500 MHz, METHANOL-d4) 88.77-8.83 (m, 1 H), 8.37-8.45 (m, 1 H), 7.54 (s, 1 H), 7. 26-7.36 (m, 1 H), 7.03 7.17 (m, 2 H), 6.93 (s, 2 H), 6.48-6.55 (m, 1 H), 6 .00 (s, 2 H), 4.60-4.75 (m, 1 H), 3.45-3.55 (m, 1 H), 2.49-2.58 (m, 1 H) , 2.38 (s, 3 H), 2.15-2.27 (m, 1 H), 1.74-1.94 (m, 3 H), 1.59-1.73 (m, 1 H).

Reference compound I-99

The title compound was prepared following general procedure B, except that methyl morpholine-3-carboxylate (4 equiv.) was the amine reagent, Hunig's base (3 equiv.) was used instead of triethylamine, and the content was heated to 120 °C for 2 h as a solution in NMP. The solvent was removed and the crude material was purified by reverse phase HPLC to provide the desired compound, reference compound I-99 (7 mg, 25% yield) as a solid.

1H NMR (400 MHz, CD3OD) 88.71-8.75 (m, 1 H), 8.21-8.26 (m, 1 H), 7.39-7.43 (m, 1 H), 7.19-7.29 (m, 1 H), 6.97-7.12 (m, 2 H), 6.85-6.88 (m, 1 H), 6.75-6.82 ( m, 1 H), 5.90-5.95 (m, 2 H), 5.20-5.31 (m, 1 H), 4.45 (s, 1 H), 3.93-4, 01 (m, 1 H), 3.82-3.93 (m, 2 H), 3.66-3.75 (m, 2 H).

Reference compound I-105

The title compound was prepared following general procedure B, except that piperidine-2-carboxamide (4 equiv.) was the amine reagent, 6 equiv. of triethylamine was used, and the contents were heated to 60 °C for 48 h. as a solution in THF/water (9:1). The solvent was removed under a stream of nitrogen, and the crude material was purified by reverse phase HPLC using a gradient of 20-51% acetonitrile/water (in 0.1% TFA) providing the desired compound, reference compound I. -105 (12 mg, 48% yield) as a solid.

1H NMR (500 MHz, METHANOL-d4) 88.80-8.87 (m, 1 H), 8.35-8.41 (m, 1 H), 7.58-7.65 (m, 1 H) ), 7.28-7.36 (m, 1 H), 7.05-7.16 (m, 2 H), 6.91-7.02 (m, 2 H), 6.03 (s, 2 H), 5.53-5.61 (m, 1 H), 4.65-4.77 (m, 1 H), 3.56-3.69 (m, 1 H), 2.37 2 .46 (m, 1 H), 1.62-2.07 (m, 6 H).

Reference compound I-106

The title compound was prepared following general procedure B, except that 4-aminotetrahydro-2 H-pyran-4-carboxylic acid (3.5 equiv.) was the amine reagent, 5 equivalents of triethylamine were used, and the Content was heated up to 200 °C for 10 min in the microwave as a solution in NMP. The reaction mixture was diluted with water and filtered. The filtrate was made alkaline to pH 10 with a 3N sodium hydroxide solution and extracted with dichloromethane. The filtrate was then acidified to pH 1 with a 1N hydrochloric acid solution and extracted with dichloromethane. The organic layer was concentrated in vacuo and the crude material was purified by reverse phase HPLC to provide the desired compound, reference compound I-106 (2.3 mg, 9% yield) as a solid.

1H NMR (500 MHz, METHANOL-d4) 88.81-8.87 (m, 1 H), 8.31-8.35 (m, 1 H), 7.38-7.41 (m, 1 H) ), 7.25-7.34 (m, 1 H), 7.04-7.15 (m, 2 H), 6.89-6.98 (m, 2 H), 6.01 (s, 2 H), 3.87-3.96 (m, 2 H), 3.76-3.87 (m, 2 H), 2.36-2.45 (m, 2 H), 2.23 2 .33 (m, 2H).

Reference compound I-110

The title compound was prepared following general procedure B, except that 4-amino-1-(tert-butoxycarbonyl)piperidine-4-carboxylic acid (3 equiv.) was the amine reagent, 5 equivalents of triethylamine were used, and the Content was heated to 120 °C for 18 h as a solution in DMSO. Without processing, the crude material was purified by reverse phase HPLC providing the desired compound, reference compound 1-110 (8.2 mg, 26% yield) as a solid.

1H NMR (500 MHz, METHANOL-d4) 88.83 (s, 1 H), 8.31-8.36 (m, 1 H), 7.39 (s, 1 H), 7.26-7, 36 (m, 1 H), 7.02-7.14 (m, 2 H), 6.91 (s, 2 H), 6.00 (s, 2 H), 3.75-3.88 ( m, 2 H), 3.36-3.49 (m, 2 H), 2.29 (br. s., 4 H), 1.50 (s, 9 H).

Reference compound I-111

The title compound was prepared following general procedure B, except that 1,2,3,4-tetrahydroisoquinoline (2.5 equiv.) was the amine reagent, no triethylamine was used, and the contents were heated to 120° C for 18 h as a solution in THF. The solvent was removed under a stream of nitrogen, and the crude material was purified by reverse phase HPLC using a gradient of 20-51% acetonitrile/water (in 0.1% TFA) providing the desired compound, reference compound I. -111 (13.9 mg, 55% yield) as a solid.

1H NMR (500 MHz, METHANOL-d4) 88.82-8.86 (m, 1 H), 8.28-8.34 (m, 1 H), 7.68-7.73 (m, 1 H) ), 7.24-7.35 (m, 6 H), 6.93-7.15 (m, 5 H), 6.00-6.06 (m, 2 H), 5.24 (s, 2 H), 4.27-4.33 (m, 2 H), 3.10-3.16 (m, 2 H).

Reference compound I-122

In a 25 ml flask, reference compound I-110 (0.056 g, 0.096 mmol) was dissolved in DCM (volume: 2 ml), and TFA (2 ml, 26.0 mmol). After stirring for 3 hours at room temperature, the reaction was complete. The solvent was removed in vacuo giving a pure product, reference compound I-122 (13.9 mg, 55% yield) as a white solid.

1H NMR (500 MHz, METHANOL-d4) 88.78-8.86 (m, 1 H), 8.30-8.38 (m, 1 H), 7.26-7.38 (m, 2 H ), 7.01-7.15 (m, 2 H), 6.84-6.96 (m, 2 H), 5.97 (s, 2 H), 3.36-3.51 (m, 4 H), 2.50-2.67 (m, 4 H).

Reference compound I-126

The title compound was prepared following general procedure B, except that 6-methoxy-1,2,3,4-tetrahydroisoquinoline-1-carboxylic acid (3 equiv.) was the amine reagent, no triethylamine was used, and the content was heated to 120 °C for 18 h as a solution in DMSO. The reaction mixture was filtered and directly purified by reverse phase HPLC to provide the desired compound, reference compound I-126 (11 mg, 38% yield) as a solid.

1H NMR (500 MHz, METHANOL-d4) 88.82-8.85 (m, 1 H), 8.33-8.37 (m, 1 H), 7.53-7.59 (m, 2 H ), 7.27-7.34 (m, 1 H), 7.10 (m, 2 H), 6.86-6.97 (m, 4 H), 5.01 (s, 2 H), 5.96 (m, 1 H), 4.35-4.45 (m, 1 H), 4.04-4.15 (m, 1 H), 3.84 (s, 3 H), 3, 04 (m, 2 H).

Reference compound I-127

The title compound was prepared following general procedure B, except that 6-hydroxy-1,2,3,4-tetrahydroisoquinoline-1-carboxylic acid (3 equiv.) was the amine reagent, no triethylamine was used, and the content was heated to 120 °C for 18 h as a solution in DMSO. The reaction mixture was filtered and directly purified by reverse phase HPLC to provide the desired compound, reference compound I-127 (5.2 mg, 18% yield) as a solid.

1H NMR (500 MHz, METHANOL-d4) 88.83-8.87 (m, 1 H), 8.37-8.42 (m, 1 H), 7.57-7.63 (m, 1 H) ), 7.42-7.48 (m, 1 H), 7.28-7.36 (m, 1 H), 7.05-7.16 (m, 2 H), 6.90-7, 00 (m, 2 H), 6.71-6.80 (m, 2 H), 6.02 (s, 2 H), 5.94-5.99 (m, 1 H), 4.42 4 .51 (m, 1 H), 3.99-4.13 (m, 1 H), 3.16-3.27 (m, 2 H), 2.94-3.02 (m, 1 H) .

Reference compound I-128

The title compound was prepared following general procedure B, except that 5-fluoro-1,2,3,4-tetrahydroisoquinoline-1-carboxylic acid (3 equiv.) was the amine reagent, no triethylamine was used, and The contents were heated to 60 °C for 18 h as a solution in DMSO, followed by heating to 120 °C for 1 h. The reaction mixture was filtered and directly purified by reverse phase HPLC to provide the desired compound, reference compound I-128 (5.7 mg, 20% yield) as a solid.

1H NMR (500 MHz, CD3OD) 88.83 (m, 1 H), 8.31 (m, 1 H), 7.68 (m, 1 H), 7.28 (m, 2 H), 7. 03 (m, 6 H), 6.02 (s, 2 H), 5.21 (s, 2 H), 4.27 (m, 2 H), 3.08 (m, 2 H).

Reference compound I-130

A solution of reference compound I-122 (as the TFA salt) in dichloromethane was treated with triethylamine (2 equiv.) and propionyl chloride (1.1 equiv.) at 25 °C. The reaction was stirred at 25 °C for 18 h. A suspension remained, as well as a content treated with 5 drops of NMP (the content becomes clear), and an additional 1 equivalent of both propionyl chloride and triethylamine. The contents were then stirred at 25 °C for 18 h. The solvent was removed in vacuo and the crude material was purified by reverse phase HPLC to provide the desired compound, reference compound I-130 (5.5 mg, 47% yield) as a solid.

1H NMR (500 MHz, METHANOL-d4) 88.82-8.86 (m, 1 H), 8.33-8.38 (m, 1 H), 7.38-7.42 (m, 1 H ), 7.28-7.36 (m, 1 H), 7.05- 7.15 (m, 2 H), 6.89-6.99 (m, 2 H), 6.00 (s, 2 H ), 4.04-4.13 (m, 1 H), 3.82-3.92 (m, 1 H), 3.55-3.64 (m, 1 H), 3.44 3.52 (m, 1 H), 2.21-2.52 (m, 7 H), 1.16 (t, 3 H).

Reference compound I-131

A solution of reference compound I-122 (as the TFA salt) in dichloromethane was treated with triethylamine (2 equiv.) and methyl carbonhydrochloride (1.1 equiv.) at 25 °C. The reaction was stirred at 25 °C for 18 h. A suspension remained, as well as a content treated with 5 drops of NMP (the content becomes clear), and an additional 1 equivalent of both methyl carbonhydrochloride and triethylamine. The contents were then stirred at 25 °C for 18 h. The solvent was removed in vacuo and the crude material was purified by reverse phase HPLC to provide the desired compound, reference compound I-131 (3.8 mg, 32% yield) as a solid.

1H NMR (500 MHz, METHANOL-d4) 88.81-8.86 (m, 1 H), 8.30-8.37 (m, 1 H), 7.39 (s, 1 H), 7. 26-7.34 (m, 1 H), 7.04 7.15 (m, 2 H), 6.93 (m, 2 H), 6.00 (s, 2 H), 3.88 (m , 2 H), 3.73 (s, 3 H), 3.42-3.52 (m, 2 H), 2.28-2.34 (br.s., 4 H).

Reference compound I-132

A solution of reference compound I-122 (as the TFA salt) in dichloromethane was treated with triethylamine (2 equiv.) and ethyl isocyanate (1.1 equiv.) at 25 °C. The reaction was stirred at 25 °C for 18 h. A suspension remained, as well as a content treated with 5 drops of NMP (the content becomes clear), and an additional 1 equivalent of both ethyl isocyanate and triethylamine. The contents were then stirred at 25 °C for 18 h. The solvent was removed in vacuo and the crude material was purified by reverse phase HPLC to provide the desired compound, reference compound I-132 (5.9 mg, 49% yield) as a solid.

1H NMR (500 MHz, METHANOL-d4) 88.83-8.86 (m, 1 H), 8.32-8.37 (m, 1 H), 7.38-7.41 (m, 1 H) ), 7.28-7.34 (m, 1 H), 7.05- 7.15 (m, 2 H), 6.89-6.99 (m, 2 H), 6.00 (s, 2 H) ), 3.74-3.83 (m, 2 H), 3.37-3.45 (m, 2 H), 3.19-3.26 (m, 2 H), 2.30 (s, 4H), 1.14 (s, 3H).

Reference compound I-153

The title compound was prepared following general procedure B, except that 2-(methylamino)benzoic acid was the amine reagent, 5 equivalents of triethylamine were used, and the contents were heated at 120 °C for 12 h as a solution. in THF. The solvent was removed in vacuo and the crude material was purified by reverse phase HPLC to provide the desired compound, reference compound I-153 (5.5 mg, 40% yield) as a solid.

1H NMR (500 MHz, METHANOL-d4) 88.80-8.85 (m, 1 H), 8.12-8.21 (m, 2 H), 7.71-7.78 (m, 1 H) ), 7.53-7.64 (m, 3 H), 7.28-7.36 (m, 1 H), 7.06-7.17 (m, 2 H), 6.91-7, 02 (m, 2 H), 6.04 (s, 2 H), 3.72 (s, 3 H).

Reference compound I-161 and reference compound I-162

The title compounds were prepared following general procedure B, except that 3-methylpiperidine-2-carboxylic acid was the amine reagent, Hunig's base (5 equiv.) was used instead of triethylamine, and the contents were heated to 120 °C for 18 h as a solution in THF/water (5:1). The solvent was removed in vacuo and the crude material was purified by reverse phase HPLC providing the desired compounds, the reference compound I-161 (cis, racemic, 5.1 mg, 20% yield) as a solid and the compound reference I-162 (trans, racemic, 1.3 mg, 5%) as a solid.1*3

1H NMR (400 MHz, CD3OD) Reference compound I-161 88.77-8.80 (m, 1 H), 8.30-8.34 (m, 1 H), 7.52-7.56 ( m, 1 H), 7.23-7.32 (m, 1 H), 6.99-7.12 (m, 2 H), 6.87-6.94 (m, 2 H), 5, 98 (s, 2 H), 5.24-5.30 (m, 1 H), 4.50-4.61 (m, 1 H), 3.72-3.83 (m, 1 H), 2.09-2.21 (m, 1 H), 1.91-2.00 (m, 1 H), 1.72-1.81 (m, 2 H), 1.48-1.62 ( m, 1 H), 1.22 (d, J=7.43 Hz, 3 H).

1H NMR (400 MHz, CD3OD) Reference compound I-16258.78-8.80(m, 1 H), 8.31-8.35 (m, 1 H), 7.55-7.58 (m , 1 H), 7.24-7.32 (m, 1 H), 7.01-7.12 (m, 2 H), 6.87-6.96 (m, 2 H), 5.99 (s, 2 H), 5.29-5.39 (m, 1 H), 3.44-3.57 (m, 1 H), 2.67-2.75 (m, 1 H), 1 .78-2.03 (m, 3 H), 1.54-1.72 (m, 2 H), 1.19 (d, 3 H).

Reference compound I-197

The title compound was prepared following general procedure B, except that 2-(piperidin-4yloxy)acetic acid was used as the amine reagent, Hunig's base (5 equiv.) instead of triethylamine, and the contents were heated to 100 °C for 18 h as a solution in THF/water (10:1). The solvent was removed in vacuo and the crude material was purified by reverse phase HPLC to provide the desired compound, reference compound I-197 (3 mg, 11% yield) as a solid.

1H NMR (500 MHz, METHANOL-d4) 58.80-8.84 (m, 1 H), 8.24-8.30 (m, 1 H), 7.57-7.64 (m, 1 H) ), 7.29-7.35 (m, 1 H), 7.05-7.16 (m, 2 H), 6.92-7.01 (m, 2 H), 6.00-6, 05 (m, 2 H), 4.28-4.36 (m, 2 H), 4.23 (s, 2 H), 3.97-4.05 (m, 2 H), 3.82 3 .89 (m, 1 H), 2.05-2.16 (m, 2 H), 1.87-1.95 (m, 2 H).

Reference compound I-214

The title compound was prepared following general procedure B, except that 4-aminobutanoic acid was the amine reagent, and the contents were stirred for 14 h. The crude material was purified by silica gel chromatography using a 0-10% methanol/dichloromethane gradient providing the desired compound, reference compound I-214 (20 mg, 57% yield) as a white solid.

1H-NMR (400 MHz, DMSO-d6) 5 12.09 (bs, 1 H), 9.08 (d, 1 H), 8.21 (d, 1 H), 8.19 (bs, 1 H) ), 7.55 (s, 1 H), 7.33-7.27 (m, 1 H), 7.21-7.18 (m, 2 H), 7.08 (ddd, 1 H), 6.82 (t, 1 H), 5.88 (s, 2 H), 3.50 (dd, 2 H), 2.30 (dd, 2 H), 1.86-1.79 (m, 2H).

Reference compound I-215

The title compound was prepared following general procedure B, except that 4-(methylamino)butanoic acid was the amine reagent. The crude material was purified by silica gel chromatography using a 0-10% methanol/dichloromethane gradient providing the desired compound, reference compound I-215 (31 mg, 81% yield) as a solid.

1H-NMR (400 MHz, DMSO-d6) 59.10 (d, 1 H), 8.35 (d, 1 H), 7.67 (s, 1 H), 7.33-7.28 (m , 1 H), 7.22-7.18 (m, 2 H), 7.08 (t, 1 H), 6.86 (t, 1 H), 5.90 (s, 2 H), 1 .88 (t, 2 H), 3.30 (d, 3 H), 2.30 (t, 2 H), 1.90-1.82 (m, 2 H).

Reference compound I-219

The title compound was prepared following general procedure B, except that N-methyl-D-valine was the amine reagent. The crude material was purified by silica gel chromatography using a gradient of 0-10% methanol/dichloromethane providing the desired compound, reference compound I-219 (17 mg, 43% yield) as a solid.

1H-NMR (400 MHz, CD3OD) 58.74 (d, 1 H), 8.17 (d, 1 H), 7.41 (s, 1 H), 7.27-7.22 (m, 1 H), 7.09-6.98 (m, 2 H), 6.87 (d, 1 H), 6.81 (t, 1 H), 5.93 (s, 2 H), 4.71 (d, 1 H), 3.31 (s, 3 H), 2.51-2.43 (m, 1 H), 1.14 (d, 3 H), 0.96 (d, 3 H) .

Reference compound I-221

The title compound was prepared following general procedure B, except that N-methyl-D-leucine was the amine reagent. The crude material was purified by silica gel chromatography using a gradient of 0-10% methanol/dichloromethane providing the desired compound, reference compound I-221 (31 mg, 76% yield) as a solid.1

1H-NMR (400 MHz, DMSO-d6) 5 12.86 (bs, 1 H), 9.07 (d, 1 H), 8.29 (d, 1 H), 7.44 (s, 1 H) ), 7.32-7.27 (m, 1 H), 7.20 7.15 (m, 1 H), 7.16 (d, 1 H), 7.08-7.05 (m, 1 H), 6.85 (t, 1 H), 5.83 (dd, 2 H), 3.12 (d, 3 H), 3.05-3.00 (m, 1 H), 1.91 1.82 (m, 1 H), 1.76-1.68 (m, 1 H), 1.51-1.47 (m, 1 H), 0.89 (d, 3 H), 0, 85 (d, 3 H).

Reference compound I-185

The title compound was prepared following general procedure B, except that 3-(trifluoromethyl)pyrrolidine-3-carboxylic acid was the amine reagent. The crude material was purified by silica gel chromatography using a 0-10% methanol/dichloromethane gradient providing the desired compound, reference compound I-185 (51 mg, 87% yield) as a solid.

<1>H -R M N (400 M H z, D M S O -d<6>) 5 9 .05 (d, 1 H), 8 .29 (d, 1 H), 7 .53 (s, 1 H), 7 .29 (q, 1 H), 7 .24 (d, 1 H), 7 .21 -7 .16 (m, 1 H), 7 .07 (t, 1 H), 6 .78 (t, 1 H), 5 .88 (s, 2 H), 4 .29 (d, 1 H), 3 .98 (d, 1 H), 3 .95 -3 .75 (m, 2 H), 2 .64 -2 .37 (m, 2 H).

I-180 REFERENCE COMPOSITION

The title compound was prepared following general procedure B, except that 2-amino-4,4,4-trifluorobutanoic acid was the amine reagent. The crude material was purified by silica gel chromatography using a 0-10% methanol/dichloromethane gradient providing the desired compound, reference compound I-180 (24 mg, 58% yield) as a solid.

1H-NMR (400 MHz, DMSO-da) 813.29 (bs, 1 H), 9.07 (d, 1 H), 8.26 (d, 1 H), 8.15-8.12 (m , 1 H), 7.39 (s, 1 H), 7.28 (q, 1 H), 7.18 (t, 1 H), 7.15 (s, 1 H), 7.07 (t , 1 H), 6.83 (t, 1 H), 5.84 (s, 2 H), 4.95-4.92 (m, 1 H), 3.03-2.94 (m, 2 H).

Reference compound I-178

The title compound was prepared following general procedure B, except that 2-amino-4-(methylsulfonyl)butanoic acid was the amine reagent. The crude material was purified by silica gel chromatography using a 0-10% methanol/dichloromethane gradient providing the desired compound, reference compound I-178 (6 mg, 14% yield) as a solid.

H1 NMR (400 MHz, CD3OD) 88.78 (d, 1 H), 8.29 (d, 1 H), 7.57 (s, 1 H), 7.29-7.23 (m, 1 H ), 7.10-7.04 (m, 1 H), 7.03 (t, 1 H), 6.91 (d, 1 H), 6.89 (t, 1 H), 5.98 ( s, 2 H), 5.24 (dd, 1 H), 3.38-3.25 (m, 1 H), 3.22-3.16 (m, 1 H), 2.29 (s, 3 H), 2.67 2.58 (m, 1 H), 2.47-2.38 (m, 1 H).

Reference compound I -72

This compound was prepared following the procedure described above for reference compound I-71, except that the reaction solvent was THF and the preparation was carried out with DCM and brine (22 mg, 31%).

1H-NMR (400 MHz, DMSO-d6) 89.06 (d, 1 H), 8.23 (d, 1 H), 7.48 (s, 1 H), 7.33-7.27 (m , 1 H), 7.23-7.17 (m, 2 H), 7.08 7.03 (m, 1 H), 6.77-6.73 (m, 1 H), 5.86 ( s, 2 H), 4.33-4.24 (m, 2 H), 4.11-4.03 (m, 2 H), 3.60-3.55 (m, 1 H), 3, 14-3.07 (m, 2 H), 2.85-2.78 (m, 2 H).

Reference compound I-103

The title compound was prepared following general procedure B, except that D-leucine was the amine reagent. The crude material was purified by silica gel chromatography using a 0-10% methanol/dichloromethane gradient providing the desired compound, reference compound I-103 (18 mg, 46% yield) as a solid.

1H-NMR (400 MHz, DMSO-d6) 812.67 (bs, 1 H), 9.07 (d, 1 H), 8.23 (d, 1 H), 8.04 (d, 1 H) , 7.39 (s, 1 H), 7.28 (dd, 1 H), 7.20-7.14 (m, 1 H), 7.14 (d, 1 H), 7.07 (t , 1 H), 6.84 (t, 1 H), 5.89-5.80 (m, 2 H), 4.74-4.64 (m, 1 H), 1.86-1.79 (m, 1 H), 1.70-1.58 (m, 2 H), 0.90 (d, 3 H), 0.67 (d, 3 H).

Reference compound I-148

The title compound was prepared following general procedure B, except that (R)-2-amino-3,3-dimethylbutanoic acid was the amine reagent. The crude material was purified by silica gel chromatography using a 0-10% methanol/dichloromethane gradient providing the desired compound, reference compound I-148 (33 mg, 83% yield) as a solid.

1H-NMR (400 MHz, DMSO-d6) 812.84 (br, s, 1 H), 9.09 (d, 1 H), 8.27 (d, 1 H), 7.43-7.27 (m, 2 H), 7.33-7.27 (m, 1 H), 7.18 (t, 1 H), 7.15 (d, 1 H), 7.08 (t, 1 H), 6 .85 (t, 1 H), 5.85 (s, 2 H), 4.58 (d, 1 H), 0.96 (s, 9 H).

Reference compound I-151

The title compound was prepared following general procedure B, except that (S)-2-amino-3,3-dimethylbutanoic acid was the amine reagent. The crude material was purified by silica gel chromatography using a gradient of 0-10% methanol/dichloromethane providing the desired compound, reference compound I-151 (22 mg, 59% yield) as a solid.1

1H-NMR (400 MHz, DMSO-d6) 812.84 (br, s, 1 H), 9.09 (d, 1 H), 8.27 (d, 1 H), 7.43-7.27 (m, 2 H), 7.33-7.27 (m, 1 H), 7.18 (t, 1 H), 7.15 (d, 1 H), 7.08 (t, 1 H), 6 .85 (t, 1 H), 5.85 (s, 2 H), 4.58 (d, 1 H), 0.96 (s, 9 H).

Reference compound I-137

The title compound was prepared following general procedure B, except that N-methyl-L-leucine was the amine reagent. The crude material was purified by silica gel chromatography using a gradient of 0-10% methanol/dichloromethane providing the desired compound, reference compound I-137 (14 mg, 36% yield) as a solid.

<1>H -R M N (400 M H z, C D<3>O D ) 8 8 .79 (d, 1 H), 8 .34 (d, 1 H), 7 .52 (s, 1 H), 7 , 27 (dd, 1 H), 7 .10-7 .01 (m, 2 H), 6 .95 -6 .90 (m, 2 H), 5 .98 (s, 2 H), 5 .57 - 5 .47 (m, 1 H), 3 .44 (d, 3 H), 2 .03 -1 .98 (m, 2 H), 1 .74-1.51 (m, 1 H), 1 , 00 (d, 3 H), 0 .98 (d, 3 H).

I-115 REFERENCE COMPOSITION

The title compound was prepared following general procedure B, except that ethyl 5,6,7,8-tetrahydro-[1.2,4]triazolo[4,3-a]pyrazine-3-carboxylate (4 equiv.) was the amine reagent, and the reaction was performed in THF. The reaction was taken up in dichloromethane and brine. The crude material was purified by silica gel chromatography using a 0-10% methanol/dichloromethane gradient providing the desired compound, reference compound I-115 (42 mg, 37% yield) as a solid.

1H-NMR (400 MHz, CDCl3) 88.47 (d, 1 H), 8.35 (d, 1 H), 7.40 (s, 1 H), 7.21-7.16 (m, 1 H), 7.01 (t, 1 H), 6.95 (t, 1 H), 6.84 (t, 1 H), 6.65 (d, 1 H), 5.98 (s, 2 H), 5.35 (s, 2 H), 4.59 (t, 2 H), 4.48 (q, 2 H), 4.30 (t, 2 H), 1.44 (t, 3 H).

Reference compound I-16

Intermediate 1 (0.030 g, 0.080 mmol) was diluted with THF (2.0 mL) and then loaded with 3-(trifluoromethyl)-5,6,7,8-tetrahydro[1.2,4]triazolo[4, 3 a]pyrazine (0.031 g, 0.161 mmol). The reaction was heated to 50 °C and stirred for 1 hour. At that time, LC/<m>S did not show product formation; therefore, at this time TEA (0.056 mL, 0.401 mmol) was added and the resulting reaction mixture was heated to 80 °C overnight. In the morning, clean reaction was detected by LC/MS. The crude reaction was concentrated and purified using SiO2 chromatography using a gradient of 0-50% (7:1 ACN/MeOH) in DCM providing the desired material as a white solid (32 mg, 72%).

1H-NMR (400 MHz, DMSO-d6) 89.08 (d, 1 H), 8.43 (d, 1 H), 7.62 (s, 1 H), 7.30 (dd, 1 H) , 7.23 (d, 1 H), 7.19 (t, 1 H), 7.07 (t, 1 H), 6.81 (t, 1 H), 5.89 (s, 2 H) , 5.24 (s, 2 H), 4.33-4.25 (m, 4 H).

Reference compound I-112

The title compound was prepared following general procedure B, except that D-serine was the amine reagent, and the reaction was taken up in THF/water. The crude material was purified by reverse phase HPLC providing the desired compound, reference compound I-112 (4 mg, 15% yield) as a solid.

1H-NMR (400 MHz, CD3OD) 88.78 (d, 1 H), 8.27 (dd, 1 H), 7.51 (s, 1 H), 7.29-7.23 (m, 1 H), 7.07 (t, 1 H), 7.02 (t, 1 H), 6.92-6.91 (m, 1 H), 6.88 (t, 1 H), 5.97 (s, 2 H), 5.13 (t, 1 H), 4.09 (d, 2 H).

Reference compound I-86

The title compound was prepared following general procedure B, except that D-valine was the amine reagent, and the reaction was taken up in THF/water. The crude material was purified by reverse phase HPLC providing the desired compound, reference compound I-86 (2 mg, 7% yield) as a solid. 1H-NMR (400 MHz, CD3OD) 88.80 (d, 1 H), 8.32 (d, 1 H), 7.57 (s, 1 H), 7.31-7.25 (m, 1 H), 7.10-7.02 (m, 2 H), 6.95 (s, 1 H), 6.95-6.91 (m, 1 H), 6.00 (s, 2 H) , 4.85 (d, 1 H), 2.45-2.36 (m, 1 H), 1.11 (d, 3 H), 1.10 (d, 3 H).

Reference compound I-88

The title compound was prepared following general procedure B, except that L-leucine was the amine reagent, and the reaction was taken up in THF/water. The crude material was purified by reverse phase HPLC to provide the desired compound, reference compound I-88 (3 mg, 10% yield) as a solid.

1H-NMR (400 MHz, DMSO-d6) 812.67 (bs, 1 H), 9.07 (d, 1 H), 8.23 (d, 1 H), 8.04 (d, 1 H) , 7.39 (s, 1 H), 7.28 (dd, 1 H), 7.20-7.14 (m, 1 H), 7.14 (d, 1 H), 7.07 (t , 1 H), 6.84 (t, 1 H), 5.89-5.80 (m, 2 H), 4.74-4.64 (m, 1 H), 1.86-1.79 (m, 1 H), 1.70-1.58 (m, 2 H), 0.90 (d, 3 H), 0.67 (d, 3 H).

Reference compound I-67

The title compound was prepared following general procedure B, except that glycine was the amine reagent, and the reaction was taken up in THF/water. The crude material was purified by reverse phase HPLC providing the desired compound, reference compound I-67 (8 mg, 33% yield) as a solid.1 1H-NMR (400 MHz, DMSO-d6) 812.72 ( bs, 1 H), 9.07 (d, 1 H), 8.25 (d, 1 H), 8.14 (bs, 1 H), 7.45 (s, 1 H), 7.32- 7.27 (m, 1 H), 7.21-7.16 (m, 1 H), 7.16 (d, 1 H), 7.07 (t, 1 H), 6.80 (t, 1H), 5.86 (s, 2H), 4.15 (d, 2H).

Reference compound I-69

The title compound was prepared following general procedure B, except that L-valine was the amine reagent, and the reaction was taken up in THF/water. The crude material was purified by reverse phase HPLC providing the desired compound, reference compound I-69 (24 mg, 66% yield) as a solid.

<1>H -R M N (400 M H z, C D<3>O D ) 8 8 .80 (d, 1 H), 8 .32 (d, 1 H), 7 .57 (s, 1 H), 7 , 31 -7 .25 (m, 1 H), 7 .10 -7 .02 (m, 2 H), 6 .95 (s, 1 H), 6 .95-6 .91 (m, 1 H), 6 .00 (s, 2 H), 4 .85 (d, 1 H), 2 .45 -2 .36 (m, 1 H), 1.11 (d, 3 H), 1.10 (d, 3H).

I-89 REFERENCE COMPOSITION

The title compound was prepared following general procedure B, except that N-methyl-L-valine was the amine reagent, and the reaction was taken up in THF/water. The crude material was purified by reverse phase HPLC providing the desired compound, reference compound I-89 (22 mg, 76% yield) as a solid.

1H-NMR (400 MHz, CD3OD) 68.74 (d, 1 H), 8.17 (d, 1 H), 7.41 (s, 1 H), 7.27-7.22 (m, 1 H), 7.09-6.98 (m, 2H), 6.87

(d, 1 H), 6.81 (t, 1 H), 5.93 (s, 2 H), 4.71 (d, 1 H), 3.31 (s, 3 H), 2.51 -2.43 (m, 1 H), 1.14 (d, 3 H), 0.96 (d, 3 H).

Reference compound I-79

The title compound was prepared following general procedure B, except that thiomorpholine 1,1-dioxide was the amine reagent, and the reaction was taken up in THF/water. The reaction was taken up in dichloromethane and brine. The crude material was purified by reverse phase HPLC to obtain the desired compound, reference compound I-79 (4 mg, 16% yield).

1H-NMR (400 MHz, CDCl3) 68.47-8.45 (m, 1 H), 8.33 (d, 1 H), 7.24 (s, 1 H), 7.19 (dd, 1 H), 7.02 (t, 1 H), 6.96 (t, 1 H),

6.84 (t, 1 H), 6.57 (d, 1 H), 5.94 (s, 2 H), 4.36 (dd, 2 H), 3.19 (dd, 2 H).

Reference compound I-68

The title compound was prepared following general procedure B, except that L-serine was the amine reagent, and the reaction was taken up in THF/water. The crude material was purified by reverse phase HPLC providing the desired compound, reference compound I-68 (12 mg, 48% yield) as a solid.

1H-NMR (400 MHz, CD3OD) 68.78 (d, 1 H), 8.27 (dd, 1 H), 7.51 (s, 1 H), 7.29-7.23 (m, 1 H), 7.07 (t, 1 H), 7.02 (t, 1 H),

6.92-6.91 (m, 1 H), 6.88 (t, 1 H), 5.97 (s, 2 H), 5.13 (t, 1 H), 4.09 (d, 2H).

Reference compound I-65

The title compound was prepared following general procedure B, except that ferf-butylamine (50 equiv.) was the amine reagent and the reaction was heated at 60 °C for 48 h as a solution in THF. The reaction was concentrated in vacuo and the crude material was purified by silica gel chromatography using a gradient of 0-30% (7:1 acetonitrile/methanol) in dichloromethane providing the desired compound, reference compound I-65 (19 mg, yield 96%) as a solid 1H-NMR (400 MHz, CDCl3) 68.45 (d, 1 H), 8.14 (d, 1 H), 7.40 (bs, 1 H), 7, twenty-one

7.16 (m, 1 H), 7.03-6.91 (m, 3 H), 6.61 (d, 1 H), 5.93 (s, 2 H), 1.58 (s, 9H).

Reference compound I-113

This compound was prepared by treating the reference compound I-115 with LO H H 2O in a solvent mixture

2:1:1 THF:MeOH:water. After decarboxylation was complete, the reaction was acidified with 1N HCl and then extracted (3 times) with dichloromethane. The organic parts were combined, dried (Na2SO4), filtered, and then concentrated. The crude material was purified by silica gel chromatography using a gradient of 0-10% MeOH in dichloromethane to provide the title compound, the reference compound.

I-113, as a white solid (5 mg, 5%).

1H-NMR (400 MHz, DMSO-d6) 69.07 (d, 1 H), 8.50 (s, 1 H), 8.41 (d, 1 H), 7.61 (s, 1 H) , 7.32-7.28 (m, 1 H), 7.24 (d, 1

H), 7.19 (t, 1 H), 7.07 (t, 1 H), 6.81 (t, 1 H), 5.89 (s, 2 H), 5.14 (s, 2 H), 4.28-4.16 (m, 4 H).

Reference compound I-174

The title compound was prepared following general procedure B, except that 3-aminopropanoic acid was the amine reagent, and the content was heated at 110 °C for 14 h as a solution in THF/water (10:1). The crude material was purified by preparative reverse phase HPLC to provide the desired compound, reference compound I-174 (17 mg, 56%) as a white solid.

1H NMR (500 MHz, CD3OD) 6 ppm 8.80 (br. s., 1 H), 8.17 (br. s., 1 H), 7.55 (s, 1 H), 7.29 ( d, 1 H), 7.01 - 7.15 (m, 2 H), 6.95 (br. s., 1 H), 6.91 (d, 1 H), 6.00 (br. s. ., 2 H), 3.96 (t, 2 H), 2.77 (t, 2 H).

Reference compound I-169

The title compound was prepared following general procedure B, except that 3-(methylamino)propanoic acid was the amine reagent, and the content was heated at 110 °C for 4 h as a solution in THF/water (10: 1). The reaction was concentrated in vacuo and the crude material was purified by reverse phase HPLC to provide the desired compound, reference compound I-169 (14 mg, 56% yield) as a solid.

1H NMR (500 MHz, CD3OD) 6 ppm 8.71 (d, 1 H), 8.17 (d, 1 H), 7.48 (s, 1 H), 7.19 (d, 1 H), 6.89 - 7.05 (m, 2 H), 6.84 (d, 2 H), 5.90 (s, 2 H), 4.05 (t, 2 H), 3.42 (d, 3H), 2.71 (t, 2H).

I-170 REFERENCE COMPOSITION

The title compound was prepared following general procedure B, except that 2-methyl-3-(methylamino)propanoic acid was the amine reagent, and the contents were heated at 110 °C for 18 h as a solution in THF/ water (10:1). The reaction was concentrated in vacuo and the crude material was purified by reverse phase HPLC to provide the desired compound, reference compound I-170 (13 mg, 51% yield) as a solid.

1H NMR (500 MHz, CD3OD) 8 ppm 8.68 (d, 1 H), 8.09 (d, 1 H), 7.38 (s, 1 H), 7.14 - 7.21 (m, 1 H), 6.90 - 7.02 (m, 2 H), 6.76-6.83 (m, 2 H), 5.87 (s, 2 H), 4.01 (dd, 1 H) ), 3.77 (dd, 1 H), 3.34 (d, 3 H), 2.92 (m, 1 H), 1.14 (d, 3 H).

Reference compound I-171

The title compound was prepared following general procedure B, except that (R)-2-(aminomethyl)-3-methylbutanoic acid was the amine reagent, and the content was heated to 110 °C for 18 h as a solution. in THF/water (10:1). The reaction was concentrated in vacuo, methanol was added, and the crude material was purified by reverse phase HPLC to provide the desired compound, reference compound I-171 (15 mg, 57% yield) as a solid.

1H NMR (500 MHz, M ETAN O L^) 8 ppm 8.86 (d, 1 H), 8.29 (d, 1 H), 7.60 (s, 1 H), 7.33 (d, 1 H), 7.06 - 7.17 (m, 2 H), 6.99 - 7.05 (m, 1 H), 6.95 (d, 1 H), 6.04 (s, 2 H) , 3.93 - 4.08 (m, 2 H), 2.71 (ddd, 1 H), 2.10 (dq, 1 H), 1.07 - 1.20 (m, 6 H).

Reference compound I-173

The title compound was prepared following general procedure B, except that (S)-2-(aminomethyl)-3-methylbutanoic acid was the amine reagent, and the content was heated to 110 °C for 18 h as a solution. in THF/water (10:1). The reaction was concentrated in vacuo and the crude material was purified by reverse phase HPLC to provide the desired compound, reference compound I-173 (18 mg, 68% yield) as a solid.

1H NMR (500 MHz, CD3OD) 8 ppm 8.84 (d, 1 H), 8.25 (d, 1 H), 7.56 (s, 1 H), 7.31 (d, 1 H), 7.04 - 7.15 (m, 2 H), 6.96 -7.01 (m, 1 H), 6.93 (d, 1 H), 6.01 (s, 2 H), 3, 91 -4.04 (m, 2 H), 2.71 (dt, 1 H), 2 .04-2.14 (m, 1 H), 1.14 (d, 3 H), 1.10 ( d, 3H).

Reference compound I-181

The title compound was prepared following general procedure B, except that (R)-3-amino-4-methylpentanoic acid was the amine reagent, and the content was heated at 100 °C for 18 h as a solution in THF /water (10:1). The reaction was concentrated in vacuo and the crude material was purified by reverse phase HPLC to provide the desired compound, reference compound I-181 (7 mg, 21% yield).

1H NMR (500 MHz, CD3OD) 8 ppm 8.85 (d, 1 H), 8.29 (d, 1 H), 7.65 (s, 1 H), 7.30 - 7.37 (m, 1 H), 7.07 - 7.16 (m, 2 H), 6.98-7.03 (m, 2 H), 6.05 (s, 2 H), 4.91 -4 .96 ( m , 1 H), 2.71 -2.86(m , 2 H), 2 .05-2.13 (m, 1 H), 1.08 (dd, 6 H). Reference compound I-182

The title compound was prepared following general procedure B, except that (S)-3-amino-4-methylpentanoic acid was the amine reagent, and the content was heated at 100 °C for 18 h as a solution in THF /water (10:1). The reaction was concentrated in vacuo and the crude material was purified by reverse phase HPLC to provide the desired compound, reference compound I-182 (7 mg, 24% yield).

1H NMR (500 MHz, CD3OD) 8 ppm 8.79 - 8.85 (m, 1 H), 8.23 - 8.28 (m, 1 H), 7.63 (d, 1 H), 7. 30 (br. s., 1 H), 7.03 -7.15 (m, 2 H), 6.94-7.02 (m, 2 H), 6.03 (br. s., 2 H ), 2.66-2.85 (m, 2 H), 2.01 -2.13 (m, 2 H), 1.00-1.10 (m, 6 H). Reference compound I-195 and reference compound I-196

The title compounds were prepared following general procedure B, except that 4-methyl-3-(methylamino)pentanoic acid was the amine reagent, and the contents were heated at 100 °C for 18 h as a solution in THF/ water (10:1). The reaction was concentrated in vacuo and the crude material was purified by reverse phase HPLC providing two compounds, reference compound I-195 (5 mg, 16% yield), and reference compound I-196 (12 mg, yield 41%).

1H NMR for Reference Compound I-195 (500 MHz, CD3OD) 8 ppm 8.82 (d, 1 H), 8.28 (d, 1 H), 7.57 (s, 1 H), 7.29 - 7.34 (m, 1 H), 7.05 - 7.15 (m, 2 H), 6.93 - 6.98 (m, 2 H), 6.02 (s, 2 H), 2, 92 (m, 2 H), 2.75 - 2.82 (m, 3 H), 2.10 -2.19 (m, 2 H), 1.13 (d, 3 H) 1.00 (d , 3H).

1H NMR for Reference Compound I-196 (500 MHz, CD3OD) 8 ppm 8.83 (d, 1 H), 8.21 (d, 1 H), 7.64 (s, 1 H), 7.29 - 7.35 (m, 1 H), 7.05 -7.14 (m, 2 H), 6.95 - 7.01 (m, 2 H), 6.03 (s, 2 H), 3, 27 (s, 3 H).

Reference compound I-202

The title composition was prepared in 3 stages:

Step 1: Synthesis of 3-((5-fluoro-2-(1-(2-fluorobenzyl)-5-(isoxazol-3-yl)-1 H-pyrazol-3-yl)pyrimidin-4-yl)amino (R)-methyl )-4-metholpentanoate

To a stirred solution of reference compound I-181 in ether/methanol (3:1) TMS-diazomethane (2 equiv.) was added slowly at 23 °C. The mixture was stirred for 30 min and the solvent was removed in vacuo. The crude material was purified by silica gel chromatography providing the desired intermediate product, 3-((5-fluoro-2-(1-(2-fluorobenzyl)-5-(isoxazol-3-yl)-1 H-pyrazole (R)-methyl -3-yl)pyrimidin-4-yl)amino)-4-methylpentanoate (58 mg, 56% yield). Step 2: Synthesis of 3-((5-fluoro-2-(1-(2-fluorobenzyl)-5-(isoxazol-3-yl)-1 H-pyrazol-3-yl)pyrimidin-4-yl)( (R)-methyl methyl)amino)-4-methylpentanoate

To a 0 °C solution of 3-((5-fluoro-2-(1-(2-fluorobenzyl)-5-(isoxazol-3-yl)-1 H-pyrazol-3-yl)pyrimidin-4- (R)-methylyl)amino)-4-methylpentanoate in DMF sodium hydride (1.2 equiv.) was added followed by iodomethane (1.1 equiv.). The mixture was stirred and heated to 23°C. The reaction was quenched with water and the layers were separated. The aqueous layer was extracted with dichloromethane, and the organic layer was dried, filtered and concentrated. The residue was collected in the next step without further purification.

Stage 3: Synthesis of reference compound I-202

To a stirred solution of 3-((5-fluoro-2-(1-(2-fluorobenzyl)-5-(isoxazol-3-yl)-1 H-pyrazol-3-yl)pyrimidin-4-yl)( (R)-methyl methyl)amino)-4-methylpentanoate in THF/water/methanol (3:1:1) sodium hydroxide (3 equiv.) was added. The contents were stirred at 23 °C for 18 h. The solvent was removed in vacuo and the crude material was purified by reverse phase HPLC to provide the reference compound I-202 (0.5 mg, 12% yield) as a solid.

1H NMR (500 MHz, CD3OD) 8 ppm 8.83 (d, 1 H), 8.29 (d, 1 H), 7.58 (s, 1 H), 7.32 (dd, 2 H), 7.06 - 7.15 (m, 1 H), 6.93 -6.99 (m, 2 H), 6.02 (s, 2 H), 2.90 (dd, 2 H), 2, 75-2.82 (m, 3 H), 2.14 (m, 2 H), 1.13 (d, 3 H), 1.00 (d, 3 H).

Reference compound I-206

The title compound was prepared following general procedure B, except that 3-amino-2,2-difluoropentanoic acid was the amine reagent, and the content was heated at 110 °C for 18 h as a solution in dioxane/water (10:1). The reaction was concentrated in vacuo, methanol was added, and the crude material was purified by reverse phase HPLC to provide the desired compound, reference compound I-206 (20 mg, 22% yield).

1H NMR (500 MHz, CD3OD) 8 ppm 8.78 (d, 1 H), 8.22 (d, 1 H), 7.61 (s, 1 H), 7.25 - 7.31 (m, 1 H), 7.07 - 7.12 (m, 1 H), 7.05 (t, 1 H), 6.96 (d, 1 H), 6.89 (t, 1 H), 6, 00 (s, 2 H), 4.35 (t, 2 H).

Reference compound I-251

The title compound was prepared following general procedure B, except that (S)-3-amino-4,4-dimethylpentanoic acid was the amine reagent, and the content was heated at 110 °C for 18 h as a solution. in THF/water (10:1). The reaction was concentrated in vacuo, methanol was added, and the crude material was purified by reverse phase HPLC to provide the desired compound, reference compound I-251 (15 mg, 44% yield).1 1H NMR (500 MHz, CD3OD) 8 ppm 8.83 (d, 1 H), 8.26 (d, 1 H), 7.63 (s, 1 H), 7.29 - 7.35 (m, 1 H), 7, 06 - 7.16 (m, 2 H), 7.02 (d, 1 H), 6.95 - 7.00 (m, 1 H), 6.04 (s, 2 H), 2.82 - 2.88 (m, 1 H), 2.72 (dd, 2 H), 1.08 (s, 9 H).

Reference compound I-266

A solution of 5,5-difluoropiperidine-2-carboxylic acid (2.5-3.0 equivalents), triethylamine (8.0-10 equivalents) and intermediate 1 was stirred in dioxane/water (2:1 ratio) at 100 °C until complete consumption of the starting material according to LC/MS, following general procedure B. The solution was poured onto 1N HCl and extracted with dichloromethane. The organic phases were dried over sodium sulfate, filtered and concentrated in vacuo. Purification by silica gel chromatography (3-8% methanol/dichloromethane gradient) afforded the desired compound, reference compound I-266, (29 mg, combined yield from 2 experiments) as an off-white solid.

1H -R M N (400 M H z, C D C h ) 8 8 .46 (d, 1 H), 8 .25 (d, 1 H), 7 .36 (s, 1 H), 7 .20 (ap. q, 1 H), 7 .03 (ap. t, 1 H), 6 .96 (ap. t, 1 H), 6 .69 (ap. t, 1 H), 6 .58 (d, 1 H), 6 .22 (d, 1 H), 6 .08 (d, 1 H), 5 .95 (m, 1 H), 4 .59 (m, 1 H), 3 .53 (dd, 1 H), 2 .37 (br. d, 1 H), 2 .08 (m, 2 H), 1.57 (m, 1 H).

REFERENCE COMPOSITION I-263

The title composition was prepared in 4 stages:

Stage 1: Synthesis of ferf-butyl 4,4-difluoropiperidin-1-carboxylate

A suspension of 4,4-difluorpiperidine hydrochloride and triethylamine (2.2 equivalents) in dichloromethane was added to a solution of di-ferf-butyl dicarbonate (1.1 equivalents) in dichloromethane slowly by pipette (note: observed gas release). The reaction was stirred at room temperature until complete consumption of the starting material as indicated by NMR. The reaction mixture was diluted with dichloromethane and washed with half-saturated ammonium chloride solution. The organic layer was dried over sodium sulfate, filtered, and the solvent was removed in vacuo. Purification by silica gel chromatography (2% ethyl acetate/hexane) afforded ferf-butyl 4,4-difluoropiperidine-1-carboxylate (73%).

Stage 2: Synthesis of 1-(ferf-butoxycarbonyl)-4,4-difluorpiperidine-2-carboxylic acid

A 0.5 M solution of ferf-butyl 4,4-difluoropiperidin-1-carboxylate, tetramethylethylenediamine (TMEDA, 1.0 equivalent) in anhydrous ether at -78 °C was treated dropwise with sec-butyllithium (1, 2 equivalents) and stirred for 2 hours. Carbon dioxide gas was then introduced by bubbling for 2 min. The reaction was stirred at −78 °C for 10 min, warmed to room temperature, and stirred for an additional hour. The resulting mixture was then quenched with water, acidified to pH 2 with 1N HCl and extracted with ethyl acetate. The combined organic layers were dried over sodium sulfate, filtered, and the solvent was removed in vacuo. Purification by silica gel chromatography (20-50% ethyl acetate/hexane gradient) afforded 1-(ferfbutoxycarbonyl)-4,4-difluoropiperidine-2-carboxylic acid (87%).

Stage 3: Synthesis of 2-carboxy-4,4-difluoropiperidinium trifluoroacetate

A solution of trifluoroacetic acid (20 equivalents) and 1-(ferf-butoxycarbonyl)-4,4-difluoropiperidine-2-carboxylic acid was stirred in dichloromethane at room temperature until complete consumption of starting material determined by LC/MS. The reaction mixture was concentrated in vacuo to provide 2-carboxy-4,4-difluoropiperidinium trifluoroacetate (WW intermediate) as a sticky pale orange solid (99%) which was used without further handling.

Stage 4: Synthesis of reference compound I-263

The title compound was prepared following general procedure B, except that 2-carboxy-4,4-difluoropiperidinium trifluoroacetate (2.4 equiv.) was the amine reagent, and the contents were heated to 100 °C. The crude material was purified by silica gel chromatography with a gradient of 2-7% methanol/dichloromethane providing the desired reference compound I-263 (26 mg, 56% yield) as an off-white solid.

1H-NMR (400 MHz, acetone-da) 88.88 (s, 1 H), 8.30 (d, 1 H), 7.50 (s, 1 H), 7.31 (ap. q, 1 H), 7.14 (ap. t, 1 H), 7.08 (ap. t, 1 H), 7.01 (s, 1 H), 6.90 (ap. t, 1 H), 5 .96 (s, 2 H), 5.70 (br. d, 1 H), 4.69 (br. d, 1 H), 3.68 (ap. t, 1 H), 2.84 (m, 1 H), 2.52 (m, 1 H), 2.23 (m, 2 H).

Reference compound I-247

A mixture of intermediate 1 (26.0 mg), (2S)-3-methyl-2-(methylamino)pentanoic acid (0.030 g, 3 equiv, and triethylamine (0.096 mL, 10 equiv) in a 10:1 mixture of THF/water was heated to 85 °C for 16 h, following procedure B. The reaction was cooled, the solvent was removed, and the resulting crude product was purified by preparative reverse phase HPLC providing the desired product, the compound reference I-247 as a solid (2.4 mg, yield 7.2 %).

d 1H NMR (500 MHz, CD3OD) 88.80 (d, 1 H), 8.32 (d, 1 H), 7.51 (s, 1 H), 7.26 - 7.32 (m, 1 H), 7.02 - 7.13 (m, 2 H), 6.92 - 6.95 (m, 1 H), 6.91 (d, 1 H), 5.99 (s, 2 H) , 5.49 (s, 1 H), 3.44 (d, 3 H), 2.03 (s, 1 H), 1.03 (t, 3 H), 0.98 (dd, 2 H) , 0.85-0.92 (m, 3 H).

I-255 REFERENCE COMPOSITION

A mixture of intermediate-i (36.0 mg), 4-isopropylpiperidine-4-carboxylic acid (3equiv.), and TEA (10 equiv.), in a 10:1 THF/water mixture was heated to 90° C for 3 hours, following general procedure B. The reaction

cooled, the solvent was removed, and the resulting crude product provided the desired product, reference compound I-255, as a white solid (22 mg, 49% yield)

1H NMR (500 MHz, CHLOROFORM-d) 6 ppm 8.49 (d, 1 H), 8.35 (d, 1 H), 7.64 (s, 1 H), 7.48 (s, 1 H) ), 7.20 - 7.25 (m,

1 H), 7.05 (s, 1 H), 7.01- 7.05 (m, 3 H), 6.67 (d, 1 H), 5.98 (s, 2 H), 4, 80 (d, 2 H), 3.72 - 3.79 (m, 1 H), 3.23 (t, 2 H), 2.35

(d, 3 H), 1.80 - 1.92 (m, 2 H), 1.62 (td, 2 H), 1.41 (t, 1 H), 0.97 (d, 6 H)

Reference compound I-254

A mixture of intermediate-1 (35.0 mg), 42-(piperidin-4-yl)benzoic acid (3 equiv.), and TEA (10 equiv.), in a 10:1 THF/water mixture was heated at 90 °C for 2 hours. The reaction mixture was cooled, the solvent was removed, and the mixture was treated with 1N HCl and the resulting crude product was purified by preparative reverse phase HPLC to provide the desired product, reference compound I-254, as a white solid. (1 mg, 2% yield).

1H NMR (500 MHz, CDCh) 6 ppm 8.45 (d, 1 H), 8.22 (d, 1 H), 7.99 (dd, 1 H), 7.51 - 7.56 (m, 1H), 7.39 - 7.44(m, 2H),

7.32 (t, 1 H), 7.17 - 7.24 (m, 1 H), 6.95 - 7.09 (m, 2 H), 6.86 - 6.93 (m, 1 H) ), 6.61 (s, 1 H), 5.98 (s, 2 H), 4.90 (br. s. H), 3.94 (br. s., 1 H), 3.21 ( t, 2H), 2.03 - 2.09 (m, 2H), 1.80 - 1.90 (m, 2H)

Reference compound I-256

A mixture of intermediate 1 (20.0 mg), 4-(tert-pentyl)piperidine-4-carboxylic acid (3 equiv, as salt of

TFA), and TEA (10 equiv.), in a 10:1 THF/water mixture was heated at 90 °C for 2 hours. The reaction was cooled, the organic solvent was removed, 1N HCl was added and the resulting precipitate was filtered to provide the desired product, reference compound I-256, as a white solid (13.4 mg, 47% yield).

1H NMR (500 MHz, CDCla) 6 ppm: 8.45 (d, 1 H), 8.17 (d, 1 H), 7.29 (s, 1 H), 7.15 - 7.22 (m , 1H), 6.99 - 7.07 (m, 1H),

6.96 (t, 1 H), 6.85 (t, 1 H), 6.58 (d, 1 H), 5.97 (s, 2 H), 4.69 (d, 2 H), 3.04 (t, 2H), 2.25 (d, 2H), 1.71 (td, 2H), 1.35

1.45 (m, 2 H), 0.90 - 0.95 (m, 6 H), 0.87 (t, 3 H)

Reference compound I-258

To a solution of (S)-2-((5-fluoro-2-(1-(2-fluorobenzyl)-5-(isoxazol-3-yl)-1 H-pyrazol-3-yl)pyrimidin-4 acid -yl)amino)-3-methylbutanoic acid (reference compound I-69, 0.040 g, 0.088 mmol) in DCM (1.8 ml) CDI (0.043 g, 0.264 mmol) was added. The reaction was heated at 45 °C for 60 minutes. After that, DBU (0.013 ml, 0.088 mmol) and cyclopropanesulfonamide (0.053 g, 0.440 mmol) were added. The reaction was continued for a further 40 minutes at the same temperature, until it was considered complete. At that time, the reaction was quenched with 1N HCl. The layers were separated and the aqueous part was extracted twice with DCM. The organic parts were combined, dried (Na2SO4), filtered, and concentrated. The crude material was purified by silica chromatography with a gradient

of 0-10% MeOH/DCM providing the desired compound, compound I-258, as a white solid (10.8 mg, 80% yield).

1H NMR (500 MHz, CHLOROFORM-d) 6 ppm: 9.93 (br. s., 1 H), 8.46 (d, 1 H), 8.21 (d,1 H), 7.32 ( s, 1H), 7.21 - 7.25

(m, 1 H), 6.99 - 7.09 (m, 2 H), 6.91 - 6.97 (m, 1 H), 6.61 (d, 1 H), 6.03 - 6 .08 (m, 1 H), 5.93 - 5.99 (m, 1 H), 5.45 (d, 1

H), 4.35 (t, 1 H), 2.77 - 2.88 (m, 1 H), 2.52 - 2.62 (m, 1 H), 1.12-1.15 (m , 6 H), 1.05 - 1.07 (m, 2 H), 0.88 - 0.90 (m, 2

H).

Reference compound I-259

To a solution of the reference compound I-88 in dichloromethane was added CDI (3 equiv.). The reaction was heated to

45 °C for 30 min, after which DBU (1 equiv.) and methanesulfonamide (5 equiv.) were added. The reaction was heated for an additional 40 min until the reaction was complete. At that time, the reaction was quenched with 1N hydrochloric acid solution. The layers were separated and the aqueous part was extracted with dichloromethane (2 times). The organic parts were combined, dried (Na2SO4), filtered, and concentrated. The crude material was purified by silica gel chromatography with a gradient of 0-10% methanol/dichloromethane providing the desired compound, reference compound I-259 (15.8 mg, 44% yield) as a white solid.1

1H NMR (500 MHz, CDCta) 6 ppm 10.14 (br. s., 1 H), 8.47 (d, 1 H), 8.22 (d, 1 H), 7.33 (s, 1 H), 7.21 - 7.26 (m, 1H),

6.99 - 7.08 (m, 2 H), 6.91 - 6.96 (m, 1 H), 6.63 (d, 1 H), 5.96- 6.09 (m, 2 H ), 5.3 (br. s., 1 H) 4.51 - 4.60 (m, 1 H), 3.06

- 3.11 (m, 3 H), 1.90-2.00 (m, 1 H), 1.71 - 1.87 (m, 2 H), 1.05 (d, 3 H), 0 .96-0.99 (m, 3 H).

Reference compound I-261

To a solution of the reference compound I-103 in dichloromethane was added CDI (3 equiv.). The reaction was heated

at 45 °C for 1.5 h, after which DBU (1 equiv.) and cyclopropanesulfonamide (5 equiv.) were added. The reaction

It was heated for a further 40 minutes, until the reaction was complete. The reaction was quenched with 1N hydrochloric acid solution, the layers were separated, and the aqueous part was extracted with dichloromethane (2 times). The organic parts were combined, dried (Na2SO4), filtered, and concentrated. The crude material was purified by silica gel chromatography using a 0-10% methanol/dichloromethane gradient providing the desired compound, reference compound I-261 (40 mg, 80% yield) as a solid.

1H NMR (500 MHz, CDCh) 8 ppm: 10.20 (br. s., 1 H), 8.47 (s., 1 H), 8.22 (br. s., 1 H), 7.35 (s, 1 H), 7.17 - 7.26 (m, 1

H), 6.91 - 7.09 (m, 3 H), 6.64 (s, 1 H), 5.92 - 6.09 (m, 2 H), 5.39 (br. s., 1H), 4.59 - 4.70 (m, 1H), 2.76 - 2.89 (m, 1H),

2.50 - 2.65 (m, 1 H), 1.95 (dt, 1 H), 1.69 - 1.86 (m, 2 H), 1.04 (d, 2 H), 0, 98 (d, 3 H), 0.85 - 0.95 (m, 3 H), 0.74 - 0.83 (m, 1 H).

Reference compound I-264

A mixture of intermediate 1 (38.8 mg), 2,2-dimethylthiomorpholine 1,1-dioxide (3 equiv.), and TEA (10 equiv.), in a 10:1 THF/water mixture was heated at 90 °C for 3 h, following general procedure B. The reaction was quenched, poured into a 1:1 mixture of 1N HCl and DCM, the extracted organic materials were combined (three times), dried, purified by silica chromatography using a 0-10% methanol/dichloromethane gradient providing the desired compound, reference compound I-264 (40 mg, yield

77%) in the form of a white solid.

1H NMR (500 MHz, CDCla) 8 ppm 8.48 (d, 8.31 (d, 1 H), 7.26 (s, 1 H), 7.22 (q, 1 H), 7.02 - 7.07 (m, 1 H), 6.99 (t,1 H), 6.86 - 6.91 (m, 1 H), 6.59 (d,1 H), 5.97 (s, 2 H), 4.41 (br.s., 2 H), 4.09 (br.s., 2 H), 3.26 (t, 2 H), 1.43 (s, 6 H).

Reference compound I-270

A mixture of intermediate-1 (313 mg), (S)-4-methyl-3-(methylamino)pentanoic acid (3 equiv.), and TEA (10 equiv.), in a 10:1 mixture of THF/ Water was heated to 85 °C for 3 hours, following procedure B. The reaction was cooled, poured into a 1:1 mixture of 1N HCl and DCM, the extracted organic substances were combined (three times), dried, purified by silica gel chromatography using a 0-10% methanol/dichloromethane gradient providing the desired compound, reference compound I-270 (56 mg, yield

14%) as a white solid.

1H NMR (500 MHz, CDCh) 8 ppm 8.48 - 8.53 (m, 1 H), 8.31 (br. s., 1 H), 7.39 (s, 1 H), 7.20 - 7.26 (m, 1 H), 7.13 (t, 1

H), 6.99-7.05 (m, 2 H), 6.66 (br. s., 1 H), 5.86-5.94 (m, 2 H), 3.19 (d, 3 H), 2.85 (dd, 1 H), 2.60-2.73 (m, 1 H), 1.93

- 2.05 (m, 1 H), 1.26 (s, 1 H), 0.93 (d, 3 H), 1.08 (d, 3 H).

Reference compound I-271

A mixture of intermediate-1 (313 mg), (1R,5S,6R)-3-azabicyclo[3.1.0]hexane-6-carboxylic acid (3 equiv.).

and TEA (10 equiv.), in a 10:1 mixture of THF/water, heated at 85 °C for 3 h, following the general procedure

B. The reaction was cooled, poured into a 1:1 mixture of 1N HCl and DCM, the extracted organic substances (three times), combined, dried, purified by silica gel chromatography using a gradient of 0- 10% methanol/dichloromethane providing the desired compound, reference compound I-271 (22 mg, yield

33%) as a white solid.

1H NMR (500 MHz, CDCta) 8 ppm: 8.49 (d, 1 H), 8.35 (d, 1 H), 7.45 (br. s., 1 H), 7.23 (td, 1H), 7.05 (d, 1H), 7.01 (d, 2

H), 6.63 (d, 1 H), 5.97 (s, 2 H), 4.32 (d, 2 H), 3.95 (d, 2 H), 2.40 (br. s ., 2 H, 1.65 (t, 1 H), 0.97 (d, 1 H).

Reference compound I-268

The title compound was prepared in 2 steps:

Stage 1: Synthesis of 2-carboxy-5,5-dimethylpiperidinium trifluoroacetate

2-Carboxy-5,5-dimethyl-piperidinium trifluoroacetate was prepared as a white solid following the procedure for the synthesis of 2-carboxy-4,4-difluoropiperidinium trifluoroacetate, as described in the preparation of reference compound I- 263, except for the use of 3,3-dimethylpiperidine hydrochloride in stage 1.

Stage 2: Synthesis of reference compound I-268

The title compound was prepared following general procedure B, except that 2-carboxy-5,5-dimethylpiperidinium trifluoroacetate was the amine reagent, and the contents were heated to 100 °C. The resulting solution was poured into water and acidified to pH 3 with a 1N aqueous hydrochloric acid solution. The resulting precipitate was collected by vacuum filtration, washed with HCl solution (pH 3) and ether to provide the desired compound, reference compound I-268 (38 mg, 62% yield) as a white solid.

1H-NMR (400 MHz, DMSO-de) 813.0 (br. s, 1 H), 9.10 (d, 1 H), 8.33 (d, 1 H), 7.50 (s, 1 H), 7.33 (ap. q, 1 H), 7.22 (m, 2 H), 7.10 (ap. t, 1 H), 6.85 (ap. t, 1 H), 5 .89 (s, 2 H), 5.38 (m, 1 H), 3.99 (m, 1 H), 3.03 (m, 1 H), 2.11 (m, 1 H), 2 .01 (m, 1 H), 1.44 (br. d, 1 H), 1.32 (td, 1 H), 0.97 (s, 3 H), 0.95 (s, 3 H) .

Reference compound I-245

A solution of methyl 2-(4-aminotetrahydro-2H-pyran-4-yl)acetate hydrochloride (3.0 equivalents), triethylamine (10 equivalents) and intermediate 1 was stirred in dioxane/water (ratio 2: 1) at 100 °C until complete consumption of the starting material by LC/MS, following general procedure B. The solution was poured into water and acidified to pH 3 with 1N HCl and extracted with dichloromethane. The organic phases were dried over sodium sulfate, filtered and concentrated in vacuo. Purification by reverse phase HPLC (gradient 20-65% acetonitrile in water with 0.1% trifluoacetic acid, 20 minutes) provided the reference compound I-245 (8.8 mg, 13%) as a white solid .

1H-NMR (400 MHz, CD3OD) 88.82 (d, 1 H), 8.31 (d, 1 H), 7.53 (s, 1 H), 7.30 (ap. q, 1 H) , 7.09 (m, 1 H), 7.05 (ap. t, 1 H), 7.01 (d, 1 H), 6.97 (ap. t, 1 H), 6.00 (s , 2 H), 3.82 (dt, 2 H), 3.74 (td, 2 H), 3.20 (s, 2 H), 2.65 (br. d, 2 H), 2.06 (m, 2H).

Reference compound I-155

The title compound was prepared following general procedure B, except that (R)-pyrrolidin-2-ylmethanol was the amine reagent, and the contents were heated at 40 °C for 18 min as a solution in THF. The reaction was diluted with ethyl acetate and washed with water and brine. The organic phase was dried over Na2SO4, filtered and concentrated in vacuo. The crude material was purified by silica gel chromatography with a gradient of 20-60% ethyl acetate/hexanes providing the desired compound, reference compound I-155 (11 mg, 44% yield) as a white solid.

1H-NMR (400 MHz, CDCh) 88.43 (d, 1 H), 8.14 (d, 1 H), 7.27 (s, 1 H), 7.19 (m, 1 H), 7 .01 (ap. t, 1 H), 6.97 (ap. t, 1 H), 6.89 (ap. t, 1 H), 6.57 (d, 1 H), 5.94 (s , 2 H), 5.29 (br. s, 1 H), 4.52 (m, 1 H), 3.88 (m, 2 H), 3.78 (m, 2 H), 2.13 (m, 1 H), 2.07-1.92 (m, 2 H), 1.79 (m, 1 H).

Reference compound I-160

The title compound was prepared following general procedure B, except that piperidin-2-ylmethanol was the amine reagent, and the contents were heated at 55 °C for 4 h as a solution in THF/DMSO (2:1). . The reaction was diluted with ethyl acetate and washed with water and brine. The organic phase was dried over Na2SO4, filtered and concentrated in vacuo. The crude material was purified by silica gel chromatography with a gradient of 30-60% ethyl acetate/hexanes providing the desired compound, reference compound I-160 (9.6 mg, 70% yield) as a clear oil. .

1H-NMR (400 MHz, CDCla) 88.44 (d, 1 H), 8.17 (d, 1 H), 7.26 (s, 1 H), 7.19 (m, 1 H), 7 .02 (ap. t, 1 H), 6.97 (ap. t, 1 H), 6.86 (ap. t, 1 H), 6.58 (d, 1 H), 5.97 (d , 1 H), 5.93 (d, 1 H), 4.81 (m, 1 H), 4.25 (m, 1 H), 4.14 (m, 1 H), 3.83 (br s, 1 H), 3.79 (m, 1 H), 3.28 (m, 1 H), 1.85-1.65 (m, 6 H).

Reference compound I-183

The title compound was prepared following general procedure B, except that detert-butyl pyrazoline-1-carboxylate (1.1 equiv.) was the amine reagent, and the contents were heated to 70 °C for 5 d as a solution in THF/DMSO (4:1). The reaction was poured into water and extracted with ethyl acetate. The organic phase was dried over Na2SO4, filtered and concentrated in vacuo. The crude material was purified by silica gel chromatography using a gradient of 20% ethyl acetate/hexane providing the desired compound, reference compound I-183 (53 mg, 75% yield).1

1H-NMR (400 MHz, CDCls) 88.44 (d, 1 H), 8.26 (d, 1 H), 7.30 (s, 1 H), 7.18 (m, 1 H), 7 .01 (ap. t, 1 H), 6.95 (ap. t, 1 H), 6.84 (ap. t, 1 H), 6.56 (d, 1 H), 5.96 (s , 2 H), 4.40-3.60 (br. m, 4 H), 2.13 (ap. quintuplet, 2 H), 1.45 (s, 9 H).

Reference compound I-193

A solution of trifluoroacetic acid (20 equivalents) and reference compound I-183 was stirred in dichloromethane at room temperature until complete consumption of the starting material as determined by LC/MS. The solution was carefully poured into saturated sodium bicarbonate and dichloromethane. The layers were separated and the organic layer was dried over sodium sulfate, filtered, and the solvent was removed in vacuo to provide the reference compound I-193 (35 mg, 85%) as a white solid.

1H-NMR (400 MHz, CDCI3) 58.43 (d, 1 H), 8.17 (d, 1 H), 7.31 (s, 1 H), 7.18 (m, 1 H), 7 .01 (ap. t, 1 H), 6.95 (ap. t, 1 H), 6.82 (ap. t, 1 H), 6.57 (d, 1 H), 5.96 (s , 2 H), 4.58 (br. s, 1 H), 3.87 (m, 2 H), 3.19 (ap. t, 2 H), 2.19 (ap. quintuplet, 2 H) .

Reference compound I-213

A solution of ethyl bromoacetate (1.0 equivalent), W,W-diisopropylethylamine (1.5 equivalent) and the reference compound I-193 was stirred in dimethylformamide at room temperature until complete consumption of starting material as determined via LC/MS. The solution was diluted with water and extracted with ethyl acetate. The combined organic layers were dried over sodium sulfate, filtered, and the solvent was removed in vacuo. Purification by silica gel chromatography (50% ethyl acetate/hexanes) gave the reference compound I-213 (20 mg, 59%) as a clear oil.

1H-NMR (400 MHz, CDCh) 58.44 (d, 1 H), 8.21 (d, 1 H), 7.32 (s, 1 H), 7.17 (m, 1 H), 7 .01 (ap. t, 1 H), 6.95 (ap. t, 1 H), 6.83 (ap. t, 1 H), 6.57 (d, 1 H), 5.96 (s, 2 H), 4.22 (q, 2 H), 3.98 (ap. t, 2 H), 3.70 (s, 2 H), 3.28 (ap. t, 2 H), 2.25 (ap. quintuplet, 2 H), 1.28 (t, 3 H).

Reference compound I-216

A solution of sodium hydroxide (3.0 N in water, 8.0 equivalents) and the reference compound I-213 was stirred in methanol at room temperature until complete consumption of the starting material as determined by LC/MS. The reaction mixture was concentrated, diluted with water and neutralized to pH 6-7 by the addition of 1N HCl. The crude product was collected by vacuum filtration and purification by reverse phase HPLC (gradient of 5-95% acetonitrile in water with 0.1% trifluoacetic acid, 20 minutes) provided the reference compound I-216 (13 mg, 72%) as a whitish solid.

1H-NMR (400 MHz, CD3OD) 59.09 (d, 1 H), 8.29 (d, 1 H), 7.53 (s, 1 H), 7.33 (m, 1 H), 7 .22 (m, 2 H), 7.10 (ap. t, 1 H), 6.84 (ap. t, 1 H), 5.90 (s, 2 H), 3.84 (m, 2 H) , 3.57 (s, 2 H), 3.12 (m, 2 H), 2.17 (ap. quintuplet, 2 H).

Reference compound I-222

A solution of 2-(pyrrolidin-2-yl)acetic acid hydrochloride (2.3 equivalents), triethylamine (10 equivalents) and intermediate 1 was stirred in dioxane/water (2:1 ratio) at 100 °C until complete consumption of starting material as determined by LC/MS following procedure B. The solution was diluted with water and neutralized to pH 3 by the addition of 1N HCl. The resulting solid was collected by filtration and dried under vacuum to provide the reference compound I-222 (63 mg, 94%).

1H-NMR (400 MHz, DMSO-de) 512.3 (s, 1 H), 9.11 (d, 1 H), 8.24 (d, 1 H), 7.48 (s, 1 H) , 7.33 (m, 1 H), 7.21 (m, 1 H), 7.11 (m, 2 H), 6.90 (m, 1 H), 5.87 (s, 2 H) , 4.62 (m, 1 H), 3.80 (m, 1 H), 3.65 (m, 1 H), 2.82 (m, 1 H), 2.39 (m, 1 H) , 2.12-1.90 (m, 3 H), 1.84 (m, 1 H).

Reference compound I-184

The compound was obtained by general procedure B, starting from intermediate product 1. Purification by silica gel chromatography (20-50% ethyl acetate/hexanes gradient) produced the reference compound I-184 (62 mg, 81 %) as a transparent oil.

1H-NMR (400 MHz, CDCla) 58.45 (d, 1 H), 8.22 (d, 1 H), 7.27 (s, 1 H), 7.19 (m, 1 H), 7 .02 (ap. t, 1 H), 6.97 (ap. t, 1 H), 6.87 (ap. t, 1 H), 6.59 (d, 1 H), 5.97 (d , 1 H), 5.93 (d, 1 H), 4.76 (br. m, 1 H), 4.34-3.96 (br. m, 3 H), 3.96-3.74 (br. m, 2 H), 3.50-3.10 (br. m, 4 H), 1.49 (s, 9 H).

Reference compound I-211 and reference compound I-212

A solution of trifluoroacetic acid (20 equivalents) and reference compound I-184 was stirred in dichloromethane at room temperature until complete consumption of the starting material as determined by LC/MS. The solution was carefully poured into saturated sodium bicarbonate and extracted with dichloromethane. The combined organic layers were dried over sodium sulfate, filtered, and the solvent was removed in vacuo. Purification by reverse phase HPLC (gradient 5-75% acetonitrile in water with 0.1% trifluoacetic acid, 20 minutes) yielded two products: Reference compound I-211 (17 mg, 28% as TFA salt) like a transparent oil.

1H-NMR (400 MHz, CD3OD) 58.79 (m, 1 H), 8.35 (m, 1 H), 7.52 (s, 1 H), 7.28 (m, 1 H), 7 .10 (m, 1 H), 7.04 (m, 1 H), 6.90 (m, 1 H), 6.85 (m, 1 H), 5.97 (s, 2 H), 5 .08 (m, 1 H), 4.91 (m, 1 H), 4.09 (m, 1 H), 4.01 (m, 1 H), 3.85 (ap. t, 1 H) , 3.71 (ap. d, 1 H), 3.52 (ap. d, 1 H), 3.45 (m, 1 H), 3.40 (m, 1 H).

Reference compound I-212 (19 mg, 32%) as a clear oil. 1H-NMR (400 MHz, CD3OD) 58.80 (s, 1 H), 8.34 (d, 1 H), 7.61 (d, 1 H), 7.29 (m, 1 H), 7 .10 (ap. t, 1 H), 7.05 (ap. t, 1 H), 6.94 (m, 1 H), 6.91 (ap. t, 1 H), 6.00 (s , 2 H), 5.12-4.98 (m, 1 H*), 4.82 (m, 1 H), 4.59-4.32 (m, 1 H*), 4.17 (m , 1 H*), 3.93-3.58 (m, 4 H*), 3.65-3.33 (m, 1 H*). Series of rotameric peaks (~0.5 H each) seen for selected protons marked *.

I-150 REFERENCE COMPOSITION

The title compound was prepared in 3 steps:

Step 1: Synthesis of 5-fluoro-2-(1-(2-fluorobenzyl)-5-(isoxazol-3-yl)-1H-pyrazol-3-yl)-6-hydroxypyrimidin-4(3H)-one ( The preparation of this compound is described in a patent application, WO2013/101830).

A mixture of 1-(2-fluorobenzyl)-5-(isoxazol-3-yl)-1H-pyrazole-3-carboximidamide, diethyl 2-fluoromalonate (1 equiv.) and DBU (1 equiv.) in ethanol was heated at 70 °C for 24 h. The mixture was concentrated in vacuo to provide an oil. The oil was purified by column chromatography (0 to 20% dichloromethane in methanol) to give 5-fluoro-2-(1-(2-fluorobenzyl)-5-(isoxazol-3-yl)-1H-pyrazole-3- yl)-6-hydroxypyrimidin-4(3H)-one (145 mg, 100% yield) as a white solid.

1H NMR (500 MHz, CD3OD) 8 ppm 8.81 (d, 1 H), 7.42 (s, 1 H), 7.26 - 7.36 (m, 1 H) 7.05 - 7.18 (m, 2 H), 6.97 (t, 1 H), 6.92 (d, 1 H), 5.97 (s, 2 H).

Step 2: Synthesis of 3-(3-(4,6-dichloro-5-fluoropyrimidin-2-yl)-1-(2-fluorobenzyl)-1H-pyrazol-5-yl)isoxazole

A mixture of 5-fluoro-2-(1-(2-fluorobenzyl)-5-(isoxazol-3-yl)-1H-pyrazol-3-yl)-6-hydroxypyrimidin-4(3H)-one (1 equiv .) and POCl3 (40 equiv.) was heated at 70 °C for 24 h. The mixture was concentrated in vacuo giving a white solid. It was diluted in ethyl acetate and washed with water. The organic layer was dried, filtered and evaporated to give 3-(3-(4,6-dichloro-5-fluoropyrimidin-2-yl)-1-(2-fluorobenzyl)-1H-pyrazol-5-yl)isoxazole ( 61 mg, yield 38%) as a white solid. 1H NMR (500 MHz, CDCh) 8 ppm 8.40 (s, 1 H) 7.37 (s, 1 H) 7.10 - 7.18 (m, 1 H) 6.88 - 7.00 (m , 2 H) 6.76 (t, 1 H) 6.53 (d, 1 H) 5.96 (s, 2 H).

Stage 3: Synthesis of reference compound I-150

A mixture of 3-(3-(4,6-dichloro-5-fluoropyrimidin-2-yl)-1-(2-fluorobenzyl)-1H-pyrazol-5-yl)isoxazole (1 equiv.), morpholine [1 M in THF] (1 equiv.) and Hunig's base (1 equiv.) in THF were stirred at 23 °C for 24 hours. The mixture was diluted in ethyl acetate and washed with 1N HCl solution. The organic layer was dried, filtered and evaporated to give a white solid. The solid was purified by column chromatography (0-100% ethyl acetate in hexanes) to provide the desired compound, reference compound I-150 (32 mg, 47% yield) as a white solid.

1H NMR (500 MHz, CDCla) 8 ppm 8.47 (d, 1 H), 7.31 - 7.34 (m, 1 H), 7.19 - 7.26 (m, 1 H), 7, 02 - 7.09 (m, 1 H), 6.99 (t, 1 H), 6.84 (t, 1 H), 6.59 (d, 1 H), 6.00 (s, 2 H) ), 3.89 - 3.96 (m, 4 H), 3.81 - 3.89 (m, 4 H).

Reference compound I-172

This compound was prepared in two stages.

Step 1: Synthesis of 3-(3-(4-chloro-5-fluoro-6-methoxypyrimidin-2-yl)-1-(2-fluorobenzyl)-1H-pyrazol-5-yl)isoxazole.

To a suspension of 3-(3-(4,6-dichloro-5-fluoropyrimidin-2-yl)-1-(2-fluorobenzyl)-1H-pyrazol-5-yl)isoxazole (1 equiv.) in methanol was added sodium methoxide [0.5 M in methanol] (1 equiv.). The mixture was stirred at 23 °C for 4 h. The mixture was treated with HCl (4.0 M in dioxane, 1 equiv.). The mixture was concentrated in vacuo. The resulting solid was dissolved in ethyl acetate and washed with brine. The organic layer was dried, filtered and evaporated to give 3-(3-(4-chloro-5-fluoro-6-methoxypyrimidin-2-yl)-1-(2-fluorobenzyl)-1H-pyrazol-5-yl) isoxazole (42 mg, 85% yield) as a white solid.1 1H NMR (500 MHz, CD3OD) 8 ppm 8.67 (d, 1 H), 7.45 (s, 1 H), 7.16 - 7 .21 (m, 1 H), 6.97 - 7.04 (m, 1 H), 6.94 (t, 1 H), 6.83 (d, 1 H), 6.74 (t, 1 H), 5.88 (s, 2 H), 4.12 (s, 3 H).

Stage 2: Synthesis of reference compound I-172

A mixture of 3-(3-(4-chloro-5-fluoro-6-methoxypyrimidin-2-yl)-1-(2-fluorobenzyl)-1H-pyrazol-5-yl)isoxazole (1 equiv.), triethylamine (2 equiv.) and morpholine (2 equiv.) in THF was stirred at 100 °C for 24 h. The mixture was diluted in ethyl acetate and washed with water. The organic layer was dried, filtered and evaporated to give an oil. The oil was purified by column chromatography (0-100% ethyl acetate in hexanes) to give a light brown solid. The solid was rinsed with methanol providing the desired compound,

Reference compound I-172 (19 mg, 41% yield) as a white solid.

1H NMR (500 MHz, DMSO-cfe) 8 ppm 9.08 (d, 1 H), 7.56 (s, 1 H), 7.29 - 7.36 (m, 1 H), 7.18 - 7.26 (m, 2 H), 7.09 (t, 1 H), 6.72 (t, 1 H), 5.92 (s, 2 H), 4.01 (s, 3 H), 3.72 (s, 8 H).

Reference compound I-23

A suspension of 2-(1-(2-fluorobenzyl)-5-(isoxazol-3-yl)-1H-pyrazol-3-yl)-5-(trifluoromethyl)pyrimidin-4(3H)-one (this intermediate described in a published patent application, WO2012/3405 A1) in phosphoryl trichloride (75 equiv.) was heated at 70 ° C for 1 h. The phosphoryl trichloride was removed under a stream of nitrogen and the resulting crude intermediate was dissolved in tetrahydrofuran. Morpholine (30 equiv.) was added and the solution was stirred at room temperature until the reaction was complete as determined by LC/MS. The solution was poured into saturated aqueous ammonium chloride solution and dichloromethane. The layers were separated and the aqueous layer was extracted with dichloromethane. The organic substances were washed with saturated aqueous sodium chloride, dried over magnesium sulfate, filtered, and the solvent was removed in vacuo. The residue was suspended in diethyl ether and then the resulting solid was removed by filtration to provide the desired compound, reference compound I-23 (6.5 mg, 69% yield) as a white solid.

1H-NMR (400 MHz, CD3OD) 88.77 (m, 1 H), 8.66 (s, 1 H), 7.57 (s, 1 H), 7.31-7.26 (m, 1 H), 7.12-7.02 (m, 2 H), 6.94 (m, 1 H), 6.84 (t, 1 H), 5.99 (s, 2 H), 3.85 -3.81 (m, 8 H).

Reference compound I-24

The title compound was synthesized according to the procedure described for reference compound I-23, except for the use of 2-(1-(2-fluorobenzyl)-5-(isoxazol-3-yl)-1H-pyrazole-3 -yl)pyrimidin-4-(3H)-one (this intermediate was described in the previous patent application publication WO2012/3405 A1) as a starting pyrimidone. The final residue was suspended in diethyl ether and then the resulting solid was removed by filtration to provide the desired compound, reference compound I-24 (42 mg, quantitative yield) as a white solid.

1H-NMR (400 MHz, CD3OD) 88.74 (m, 1H), 8.21 (d, 1H), 7.46 (s, 1H), 7.27-7.22 (m, 1H), 7 .10-6.99 (m, 2H), 6.89 (m, 1H), 6.79 (t, 1H), 6.70 (d, 1H), 5.95 (s, 2H), 3, 77 (br s, 8H).

Reference compound I-28

The title compound was synthesized according to the procedure described for reference compound I-23, except for the use of 5-chloro-2-(1-(2-fluorobenzyl)-5-(isoxazol-3-yl)-1H -pyrazol-3-yl)pyrimidin-4(3H)-one (this intermediate is described in published patent application WO2012/3405 A1) as the starting pyrimidone. The final residue was suspended in diethyl ether and then the resulting solid was filtered to provide the desired compound, reference compound I-28 (7.5 mg, 32% yield) as a solid.

1H-NMR (400 MHz, CDCl3) 88.44 (m, 1H), 8.38 (s, 1H), 7.30 (s, 1H), 7.21-7.15 (m, 1H), 7 .03-6.98 (s, 1H), 6.95 (t, 1H), 6.82 (t, 1H), 6.57 (m, 1H), 5.95 (s, 2H), 3, 85-3.81 (m, 8H).

Reference compound I-29

The title compound was synthesized according to the procedure described for reference compound I-23, except for the use of 2-(1-(2-fluorobenzyl)-5-(isoxazol-3-yl)-1H-pyrazole-3 -yl)-6-oxo-1,6-dihydropyrimidin-5-carbonitrile (this intermediate was described in a previous patent application publication, WO2012/3405 A1) as the starting pyrimidone. The final residue was suspended in diethyl ether and then the resulting solid was removed by filtration to provide the desired compound, reference compound I-29 (18 mg, 84% yield) as a white solid.

1H-NMR (400 MHz, DMSO-d6) 89.11 (m, 1H), 8.78 (s, 1H), 7.71 (s, 1H), 7.36-7.31 (m, 1H) , 7.27-7.20 (m, 2H), 7.11 (t, 1H), 6.83 (t, 1H), 5.93 (s, 2H), 4.02-4.00 (m , 4H), 3.76-3.74 (m, 4H).

Reference compound I-73

[0595] The title compound was prepared in 2 steps:

Step 1: Synthesis of 2-(1-(2-fluorobenzyl)-5-(isoxazol-3-yl)-1H-pyrazol-3-yl)-5-methoxypyrimidin-4(3H)-one

A solution of 1-(2-fluorobenzyl)-5-(isoxazol-3-yl)-1H-pyrazole-3-carboximidamide (1 equiv.), methyl 3-(dimethylamino)-2-methoxyacrylate (3.1 equiv. .) and 1,8-diazabicycloundec-7-ene (2 equiv.) was stirred at 100 °C for 6 h. The reaction solution was diluted with dichloromethane and saturated ammonium chloride. The layers were separated and the aqueous layer was extracted with dichloromethane (2 times). The organic substances were washed with saturated ammonium chloride and brine, dried over magnesium sulfate, filtered, and the solvent was removed in vacuo. Purification of the crude material by silica gel chromatography (0-5% methanol in dichloromethane gradient) afforded 2-(1-(2-fluorobenzyl)-5-(isoxazol-3-yl)-1H-pyrazole-3- yl)-5-methoxypyrimidin-4-ol (49 mg, 35% yield) as a white solid.

1H-NMR (400 MHz, CD3OD) 88.77 (d, 1 H), 7.58 (s, 1 H), 7.38 (s, 1 H), 7.29-7.24 (m, 1 H), 7.10-7.02 (m, 2 H), 6.92 6.85 (m, 2 H), 5.96 (s, 2 H), 3.87 (s, 3 H).

Stage 2: Synthesis of reference compound I-73

The title compound was synthesized according to the procedure described for reference compound I-23, except for the use of 2-(1-(2-fluorobenzyl)-5-(isoxazol-3-yl)-1H-pyrazole-3 -yl)-5-methoxypyrimidin-4(3H)-one as the starting pyrimidone. The final residue was suspended in diethyl ether and then the resulting solid was removed by filtration to provide the desired compound, reference compound I-73 (50 mg, 86% yield) as a white solid.

<1>H -R M N (400 M Hz, D M S O -d a) 5 9 .08 (m, 1 H), 8.11 (s, 1 H), 7.51 (s, 1 H), 7 .35 - 7 .29 (m, 1 H), 7 .24 -7 .20 (m, 2 H), 7 .10 (t, 1 H), 6 .79 (t, 1 H), 5 .89 (s, 2 H), 3 .88 (s, 3 H), 3 .75-3 .71 (m, 8 H).

Reference compound I-77

A solution of piperidine-4-carboxylic acid (3 equivalents), triethylamine (10 equivalents), and intermediate 1 was stirred in tetrahydrofuran and water (1:1 ratio) at 100 °C until complete consumption of the determined starting material. according to LC/MS, following general procedure B. The same solution was diluted with concentrated 1N hydrochloric acid and ethyl acetate. The layers were separated and the aqueous layer was extracted with ethyl acetate and 5:1 dichloromethane/isopropyl alcohol. The organic substances were combined, dried over magnesium sulfate, filtered, and the solvent was removed in vacuo. Purification by reverse phase HPLC (gradient 5-75% acetonitrile in water with 0.1% triefluoacetic acid, 20 minutes) afforded the reference compound I-77 (11 mg, 44% yield) as a white solid.

1H-NMR (400 MHz, CD3OD) 58.79 (m, 1H), 8.23 (d, 1H), 7.57 (m, 1H), 7.31-7.26 (m, 1H), 7 .12-7.03 (m, 2H), 6.96 (m, 1H), 6.90 (t, 1H), 5.99 (s, 2H), 4.70 (d, 2H), 3, 51-3.45(m, 2H), 2.79-2.74(m, 1H), 2.15-2.11 (m, 2H), 1.90-1.80(m, 2H).

Reference compound I-78

The title compound was prepared following general procedure B, except that piperidine-3-carboxylic acid was the amine reagent, and the contents were heated at 90 °C for 1.5 h as a solution in THF/water (1 :1). The solution was diluted with 1N hydrochloric acid solution and ethyl acetate. The layers were separated and the aqueous layer was extracted with ethyl acetate and 5:1 dichloromethane/isopropyl alcohol. The organic substances were combined, dried over magnesium sulfate, filtered, and the solvent was removed in vacuo. The crude material was purified by reverse phase HPLC (gradient of 5-75% acetonitrile in water with 0.1% triefluoacetic acid, 20 minutes) to provide the desired compound, reference compound I-78 (7 mg, yield 28 %) as a white solid.

1H-NMR (400 MHz, CD3OD) 58.81 (m, 1H), 8.28 (d, 1H), 7.60 (m, 1H), 7.33-7.27 (m, 1H), 7 .13-7.04 (m, 2H), 6.97 6.92 (m, 2H), 6.01 (s, 2H), 4.52 (d, 1H), 4.31-4.26 ( m, 1H), 4.00 (dd, 1H), 3.93-3.84 (m, 1H), 2.84-2.78 (m, 1H), 2.23 2.12 (m, 1H) ), 2.04-1.88 (m, 2H), 1.82-1.73 (m, 1H).

Reference compound I-76

The title compound was prepared following general procedure B, except that pyrrolidine-3-carboxylic acid was the amine reagent, and the content was heated to 60 °C for 2.5 h as a solution in THF/water (1 :1). The solution was diluted with 1N hydrochloric acid solution and ethyl acetate. The layers were separated and the aqueous layer was extracted with ethyl acetate and 5:1 dichloromethane/isopropyl alcohol. The organic substances were combined, dried over magnesium sulfate, filtered, and the solvent was removed in vacuo. The crude material was purified by reverse phase HPLC (gradient of 5-75% acetonitrile in water with 0.1% triefluoacetic acid, 20 minutes) to provide the desired compound, reference compound I-76 (11 mg, yield 45 %) as a white solid.

1H-NMR (400 MHz, CD3OD) 58.83 (m, 1H), 8.29-8.27 (m, 1H), 7.63 (s, 1H), 7.33-7.29 (m, 1H), 7.13-7.05 (m, 2H), 6.99-6.96 (m, 2H), 6.03 (s, 2H), 4.23-4.03 (m, 5H) , 2.34 (br s, 2H).

Reference compound I-92

The title compound was prepared following general procedure B, except that 2-azabicyclo[2.2.1]heptane-3-carboxylic acid was the amine reagent, and the contents were heated at 90 °C for 16 h as a solution. in THF/water (10:1). After the reaction was completed, 3N hydrochloric acid solution was added and the solvent was removed in vacuo. The crude residue was taken up in water and the solid was filtered and rinsed with water to give the desired compound, reference compound I-92 (12 mg, 47% yield) as a brown solid.

1H-NMR (400 MHz, CD3OD) exists as >3 diastereomers5 8.80 (m, 1H), 8.25-8.16 (m, 1H), 7.32-7.28 (m, 2H), 7 .12-7.03 (m, 2H), 6.97-6.83 (m, 2H), 6.00-5.96 (m, 2H), 4.51-4.36 (m, 1H) , 2.99-2.90 (m, 1H), 2.25 (d, 1H), 1.94-1.57 (m, 6H).

Reference compound I-100

Sodium hydride (1.0 equivalent) and ethyl 1H-pyrrole-2-carboxylate (1 equivalent) were dissolved in tetrahydrofuran. After stirring for 15 minutes, 0.5 equivalent of pyrrolidine anion was added to a solution of intermediate 1 (1.0 equivalent) in tetrahydrofuran at room temperature. After 5 minutes, an additional 0.5 equivalent of anion was added. The solution was stirred at room temperature for 45 minutes, 45°C for 2 hours, and then at 65°C for 15 hours. An additional 1 equivalent of pyrrole anion was added, and after 1.5 hours at 65°C the solution was diluted with saturated aqueous ammonium chloride and dichloromethane. The layers were separated and the aqueous layer was extracted with dichloromethane. The organic substances were dried over magnesium sulfate, filtered, and the solvent was removed in vacuo. Purification by silica gel chromatography (0-50% ethyl acetate in hexanes) provided the reference compound I-100 (16 mg, 42% yield) as a white solid.

<1>H -R M N (400 M Hz, C D C I3 ) 5 8.71 (m, 1H), 8 .45 (m, 1H), 7 .40 (s, 1H), 7 .33 -7 .32 ( m, 1H), 7 .22 -7 .14 (m , 2H), 7 .04 -6 .95 (m, 2H), 6 .84 (t, 1H), 6 .57 (m, 1H), 6 .43 -6 .41 (m, 1H), 6 .00 (s, 2H), 4 .23 (q, 2H), 1 .25 (t, 3H).

Reference compound I-104

A solution of reference compound I-100 (1 equivalent) in tetrahydrofuran, methanol and water (3:1:1 ratio) was treated with hydrated lithium hydroxide (1.5 equivalent). The solution was stirred at room temperature for 45 minutes and at 45 °C for 3 minutes. The solution was diluted with ethyl acetate, aqueous 1N sodium hydroxide, and water. The layers were separated and the aqueous layer was acidified with aqueous 1N hydrochloric acid to pH ~ 1. The acidified aqueous layer was extracted with ethyl acetate, dried over magnesium sulfate, filtered, and the solvent was removed in vacuo. Purification by reverse phase HPLC (gradient 5-75% acetonitrile in water with 0.1% trifluoacetic acid, 20 minutes) provided the reference compound I-104 (1 mg, 6% yield) as a white solid.

1H-MRI (400 MHz, CD3OD) 58.83 (m, 1H), 8.77 (m, 1H), 7.59 (s, 1H), 7.47 (m, 1H), 7.31-7 .25 (m, 1H), 7.16-7.03 (m, 3H), 6.94 (m, 1H), 6.87 (t, 1H), 6.48-6.46 (m, 1H ), 6.00 (s, 2H).

Reference compound I-119

The title compound was prepared following general procedure B, except that 1,2,3,4-tetrahydroisoquinoline-3-carboxylic acid (as the HCl salt) was the amine reagent, and the reaction was heated as a solution in THF/water (10:1) for 16 h. The solvent was removed in vacuo and the residue was taken up in water. The solid was removed by filtration and the filtrate was purified by reverse phase HPLC (gradient of 5-75% acetonitrile in water with 0.1% trifluoacetic acid, 20 minutes) providing the desired compound, reference compound I-119 (20 mg, yield 48%) as a white solid.

1H-NMR (400 MHz, CD3OD) 58.80 (m, 1H), 8.33 (d, 1H), 7.55 (br s, 1H), 7.30-7.27 (m, 5H), 7.12-7.03 (m, 2H), 6.93 (s, 1H), 6.89 (t, 1H), 5.99 (s, 2H), 5.58 (brs, 1H), 5 .21 (d, 1H), 5.10 (d, 1H), 3.42-3.40 (m, 2H).

Reference compound I-140

This was synthesized according to general procedure B using 1,2,3,4-tetrahydroquinoline-2-carboxylic acid and a 10:1 ratio of tetrahydrofuran:water as solvent. After consumption of starting material, the solvent was removed in vacuo and the resulting residue was purified by reverse phase HPLC (gradient of 5-75% acetonitrile in water with 0.1% TFA, 20 minutes) providing the compound of reference I-140 (9 mg, 22% yield) as an orange solid.

1H-NMR (400 MHz, CD3OD) 58.79 (m, 1H), 8.35 (d, 1H), 7.47 (s, 1H), 7.30-7.19 (m, 3H), 7 .16-7.02 (m, 4H), 6.90-6.85 (m, 2H), 5.97 (s, 2H), 5.04 (t, 1H), 2.85-2.81 (m, 1H), 2.75-2.69 (m, 2H), 1.88-1.80 (m, 1H).

Reference compound I-120

The title compound was prepared following general procedure B, except that (S)-1,2,3,4-tetrahydroisoquinoline-1-carboxylic acid was the amine reagent, and the reaction was heated at 80 °C for 2 h as a solution in THF/water (10:1). After consumption of starting material, the solvent was removed in vacuo and the resulting residue was taken up in methanol, the solids were removed by filtration, the filtrate was purified by reverse phase HPLC (gradient of 5-75% acetonitrile in water with TFA at 0.1%, 20 minutes) providing the desired compound, reference compound I-120 (27 mg, 65% yield) as a white solid.

1H-NMR (400 MHz, CD3OD) 58.80 (m, 1H), 8.34 (d, 1H), 7.67-7.64 (m, 1H), 7.55 (s, 1H), 7 .34-7.26 (m, 4H), 7.12-7.03 (m, 2H), 6.91 (t, 1H), 6.87 (s, 1H), 6.02 (s, 1H) ), 5.98 (s, 2H), 4.45-3.37 (m, 1H), 4.13-4.04 (m, 1H), 3.26-3.20 (m, 1H), 3.05 (dt, 1H).

Reference compound I-121

The title compound was prepared following general procedure B, except that (R)-1,2,3,4-tetrahydroisoquinoline-1-carboxylic acid was the amine reagent, and the reaction was heated at 80 °C for 2 h as a solution in THF/water (10:1). After consumption of starting material, the solvent was removed in vacuo and the resulting residue was taken up in methanol, the solids were removed by filtration, the filtrate was purified by reverse phase HPLC (gradient of 5-75% acetonitrile in water with TFA at 0.1%, 20 minutes) providing the desired compound, reference compound I-121 (17 mg, 62% yield) as a white solid.

1H-NMR (400 MHz, CD3OD) 58.80 (m, 1H), 8.34 (d, 1H), 7.67-7.64 (m, 1H), 7.55 (s, 1H), 7 .34-7.26 (m, 4H), 7.12-7.03 (m, 2H), 6.91 (t, 1H), 6.87 (s, 1H), 6.02 (s, 1H) ), 5.98 (s, 2H), 4.45-3.37 (m, 1H), 4.13-4.04 (m, 1H), 3.26-3.20 (m, 1H), 3.05 (dt, 1H).

Reference compound I-123

The title compound was prepared following general procedure B, except that piperidine-4-carbonitrile was the amine reagent, and the reaction was heated at 80 °C for 2 h as a solution in THF/water (10:1). . After consumption of starting material, the reaction solution was diluted with dichloromethane and the solvent was dried with magnesium sulfate. After filtering and removing the solvent in vacuo, the crude material was purified by silica gel chromatography (0-70% ethyl acetate in hexanes) to provide the desired compound, reference compound I-123 (28 mg, yield 94%) as a white solid.

1H-NMR (400 MHz, CD3OD) 88.75 (m, 1H), 8.16 (d, 1H), 7.42 (s, 1H), 7.29-7.24 (m, 1H), 7 .11-7.06 (m, 1H), 7.02 (t, 1H), 6.90 (m, 1H), 6.80 (t, 1H), 5.95 (s, 2H), 4, 24-4.18 (m, 2H), 3.76-3.70 (m, 2H), 3.13 (tt, 1H), 2.08 (ddd, 2H), 1.96 1.87 (m , 2H).

Reference compound I-141

The title compound was prepared following general procedure B, except that 2-aminoacetonitrile (as the HCl salt) was the amine reagent, and the reaction was heated to 90 °C as a solution in THF/water (10: 1). After stirring for 15 h, two additional equivalents of 2-aminoacetonitrile (as the HCl salt) were added and the solution was stirred for an additional 24 h, at which time the solution was diluted with water and ethyl acetate. The layers were separated and the organic layer was washed with water. The organic substances were dried over magnesium sulfate, filtered, and the solvent was removed in vacuo. Methanol was added, and the resulting desired solid product was separated by filtration. The methanol filtrate contained dissolved product and was purified by reverse phase HPLC (gradient 5-75% acetonitrile in water, 0.1% trifluoacetic acid, 20 minutes) providing additional product, which was combined with the filtrate solids to provide the desired compound, reference compound I-141 (11 mg, 35% yield) as a tan solid.

1H-NMR (400 MHz, DMSO-de) 89.11 (m, 1H), 8.43 (t, 1H), 8.36 (d, 1H), 7.61 (s, 1H), 7.36 -7.30 (m, 1H), 7.25-7.20 (m, 2H), 7.11 (t, 1H), 6.86 (t, 1H), 5.90 (s, 2H), 4.55 (d, 2H).

Reference compound I-145

A suspension of sodium azide (1 equivalent), ammonium chloride (1 equivalent), and reference compound I-141 (1 equivalent) in W,A/-dimethylformamide was heated to 80 °C for 1 hour and then to 110 °C for 16 hours. Additional ammonium chloride and sodium azide were added and after 20 hours the solution was diluted with methanol and water. Purification by reverse phase HPLC (gradient 5-75% acetonitrile in water with 0.1% trifluoacetic acid, 20 minutes) provided the reference compound I-145 (6 mg, 43%) as a yellow solid.

1H-NMR (400 MHz, DMSO- d6) 89.10 (m, 1H), 8.37 (d, 1H), 7.59 (s, 1H), 7.35-7.30 (m, 1H) , 7.25-7.20 (m, 2H), 7.10 (t, 1H), 6.81 (t, 1H), 5.90 (s, 2H), 3.93-3.91 (m , 4H), 3.28-3.26 (m, 4H), 2.91 (br s, 2H).

Reference compound I-139

To a 0 °C solution of 2-methylpropan-2-ol (165 equivalents) in dichloromethane was added sulfuroisocyanatide chloride (150 equivalents). The solution was kept at 0°C for 20 minutes, and then 3 equivalents of the resulting sulfonyl chloride were added to a room temperature solution of reference compound I-4 (1 equivalent) and triethylamine (3 equivalents) in dichloromethane. . The solution was kept at room temperature for 30 minutes, at which time the solution was diluted with saturated aqueous sodium bicarbonate and ethyl acetate. The layers were separated and the aqueous layer was extracted with ethyl acetate. The organic substances were combined, dried over magnesium sulfate, filtered, and the solvent was removed in vacuo. Purification by silica gel chromatography (hexanes in ethyl acetate) gave the reference compound I-139 (8 mg, quantitative yield) as a tan solid.

1H-NMR (400 MHz, CD3OD) 88.76 (m, 1H), 8.21 (d, 1H), 7.46 (s, 1H), 7.29-7.24 (m, 1H), 7 .12-7.07 (m, 1H), 7.02 (t, 1H), 6.91 (m, 1H), 6.80 (t, 1H), 5.96 (s, 2H), 4, 02-3.99 (m, 4H), 3.49-3.47 (m, 2H), 1.46-1.43 (m, 9H).

Reference compound I-125

To a solution of reference compound I-4 (1 equivalent) in dichloromethane was added triethylamine (2 equivalents) followed by methanesulfonyl chloride (1.5 equivalents). The solution was stirred at room temperature for 15 minutes and then diluted with water and ethyl acetate. The layers were separated and the organic layer was washed with water. The organics were dried over magnesium sulfate, filtered, and the solvent was removed in vacuo to provide the reference compound I-125 (8.5 mg, quantitative yield) as a tan solid.

1H-NMR (400 MHz, DMSO-d6) 89.06 (m, 1H), 8.34 (d, 1H), 7.56 (s, 1H), 7.32-7.27 (m, 1H) , 7.22-7.17 (m, 2H), 7.07 (t, 1H), 6.78 (t, 1H), 5.87 (s, 2H), 3.90-3.88 (m , 4H), 3.25-3.22 (m, 4H), 2.88 (s, 3H).

Reference compound I-146

The title compound was prepared following general procedure B, (1H-tetrazol-5-yl)methanamine (as the HCl salt) was the amine reagent, and the reaction was heated at 90 °C for 3 hours as a solution in dioxane/water (3:1). After processing, the organic substances were washed with water, dried over magnesium sulfate, filtered, and the solvent was removed in vacuo to provide the desired compound, reference compound I-146 (21 mg, 60% yield) as a white solid.

<1>H -R M N (400 M Hz, C D 3O D ) 5 8.81 (m, 1H), 8 .34 (d, 1H), 7 .45 (s, 1H), 7 .32 -7 .26 (m, 1H), 7 .12 -7 .03 (m, 2H), 6 .94 -6 .90 (m, 2H), 6 .00 (s, 2H), 5 .23 (s, 2H).

Reference compound I-147

A solution of reference compound I-139 (1 equiv.) in dichloromethane and trifluoroacetic acid (200 equiv.) was stirred at 23 °C for 2 h, at which time the solvent was removed in vacuo and the residue was taken up in methanol. The solid product was removed by filtration. Purification of the filtrate by reverse phase HPLC (gradient of 5-75% acetonitrile in water with 0.1% trifluoacetic acid, 20 minutes) and combination with the previously filtered product provided the desired compound, reference compound I- 147 (2.4 mg, 41% yield) as a white solid.

1H-NMR (400 MHz, DMSO-de) 59.10 (m, 1H), 8.37 (d, 1H), 7.59 (s, 1H), 7.36-7.30 (m, 1H) , 7.24-7.20 (m, 2H), 7.10 (t, 1H), 6.88 (s, 2H), 6.81 (t, 1H), 5.90 (s, 2H), 3.91-3.89 (m, 4H), 3.11-3.09 (m, 4H).

Reference compound I-149

The title compound was prepared following general procedure B, isoindoline-1-carboxylic acid was the amine reagent, and the reaction was heated at 80 °C for 1.5 h as a solution in THF/water (10:1 ). Upon completion, as determined by LC/MS, the solvent was removed under a stream of nitrogen and the resulting solid was taken up in methanol and water (5:1). The resulting solids were removed by filtration and the filtrate was purified by reverse phase HPLC (gradient of 5-75% acetonitrile in water with 0.1% trifluoacetic acid, 20 minutes) providing the desired compound, reference compound I-149 ( 14.5 mg, yield 54%) as a tan solid.

1H-NMR (400 MHz, DMSO-de) 59.13 (br s, 1H), 8.39 (br s, 1H), 7.51 (brs, 2H), 7.45-7.39 (m, 2H), 7.36-7.31 (m, 1H), 7.27-7.21 (m, 1H), 7.14-7.05 (m, 2H), 6.96-6.75 ( m, 2H), 6.03-5.82 (m, 3H), 5.23-5.07 (m, 2H).

Reference compound I-158

To a solution of reference compound I-139 (1 equiv) in methanol was added (diazomethyl)trimethylsilane (~15 equiv) until the yellow color persisted. The solvent was removed under a stream of nitrogen providing an intermediate product that was not isolated. Dichloromethane and trifluoacetic acid (200 equivalents) were added, and after stirring at room temperature for 2 hours, the solvent was removed under a stream of nitrogen. Purification by silica gel chromatography (0-5% methanol in dichloromethane) provided the reference compound I-158 (2.1 mg, 31% yield) as a white solid.

1H-NMR (400 MHz, CD3OD) 58.74 (m, 1H), 8.19 (d, 1H), 7.44 (s, 1H), 7.28-7.22 (m, 1H), 7 .10-7.05 (m, 1H), 7.01 (t, 1H), 6.90 (m, 1H), 6.78 (t, 1H), 5.94 (s, 2H), 4, 01-3.98 (m, 4H), 2.66 (s, 3H).

Reference compound I-177

To a 0 °C solution of reference compound I-147 (1 equivalent) and pyridine (100 equivalents) in dichloromethane was added acetyl chloride (5 equivalents). After five minutes, the solution was warmed to room temperature and kept at that temperature for 1 hour. Additional acetyl chloride (5 equiv) was added and stirred at room temperature for 2.5 hours. The solution was then poured into saturated aqueous ammonium chloride and dichloromethane. The layers were separated and the aqueous layer was extracted with dichloromethane. The organics were dried over magnesium sulfate, filtered, and the solvent was removed in vacuo. Purification by reverse phase HPLC (gradient of 5-75% acetonitrile in water with 0.1% trifluoacetic acid, 20 minutes) provided the reference compound I-177 (7.9 mg, 29% yield) as a yellow solid.

1H-NMR (400 MHz, CD3OD) 58.83 (m, 1H), 8.36 (d, 1H), 7.67 (s, 1H), 7.33-7.28 (m, 1H), 7 .13-7.04 (m, 2H), 6.97-6.92 (m, 2H), 6.02 (s, 2H), 4.22-4.20 (m, 4H), 3.60 -3.58 (m, 4H), 2.03 (s, 3H).

Reference compound I-175

The title compound was prepared following general procedure B, 4-(methylsulfonyl)piperidine was the amine reagent, and the reaction was heated at 80 °C for 1.5 h as a solution in THF. After completion of the reaction, as determined by LC/MS, the solution was diluted with ethyl acetate and saturated aqueous ammonium chloride. The layers were separated and the organic layer was washed with additional ammonium chloride solution, dried over magnesium sulfate, filtered, and the solvent was removed in vacuo to provide the desired compound, reference compound I-175 (25 mg , yield 93%) as a yellow solid.

1H-NMR (400 MHz, CDCh) 58.44 (m, 1H), 8.21 (d, 1H), 7.26 (s, 1H), 7.20-7.15 (m, 1H), 7 .03-6.96 (m, 1H), 6.95 (t, 1H), 6.82 (t, 1H), 6.57 (m, 1H), 5.95 (s, 2H), 4, 83 (d, 2H), 3.18-3.05 (m, 3H), 3.07 (s, 3H), 2.26 (d, 2H), 1.93 (qd, 2H).

Reference compound I-192

The title compound was prepared following general procedure B, W-methyl-1-(1H-tetrazol-5-yl)methanamine (as HCl salt) was the amine reagent, and the reaction was heated to 90° C for 1.5 hours as a solution in dioxane/water (3:1). After completion of the reaction, as determined by LC/MS, the solution was diluted with 1 N hydrochloric acid solution and ethyl acetate. The layers were separated and the aqueous layer was extracted with dichloromethane. The organic substances were washed with water, dried over magnesium sulfate, filtered, and the solvent was removed in vacuo to provide the desired compound, reference compound I-192 (30 mg, 83%) as a white solid.

1H-NMR (400 MHz, DMSO-de) 89.11 (m, 1H), 8.34 (d, 1H), 7.44 (s, 1H), 7.35-7.30 (m, 1H) , 7.23-7.17 (m, 2H), 7.10 (t, 1H), 6.84 (t, 1H), 5.88 (s, 2H), 5.21 (s, 2H), 3.35 (m, 3H).

Reference compound I-201

The title compound was prepared following general procedure B, ethyl 2-((cyclopropylmethyl)amino)acetate was the amine reagent, and the reaction was heated at 90 °C for 16 h as a solution in dioxane. The crude material was purified by silica gel chromatography (0-70% ethyl acetate in hexanes) providing the desired compound, reference compound I-201 (32 mg, 81% yield) as a clear oil.

1H-NMR (400 MHz, CDCh) 88.43 (m, 1H), 8.17 (d, 1H), 7.21 (s, 1H), 7.19-7.13 (m, 1H), 7 .01-6.92 (m, 2H), 6.81 (t, 1H), 6.54 (m, 1H), 5.92 (s, 2H), 4.37 (s, 2H), 4, 17 (q, 2H), 3.63 (d, 2H), 1.21 (t, 3H), 1.15-1.09 (m, 1H), 0.59-0.54 (m, 2H) , 0.31-0.26 (m, 2H).

Reference compound I-203

A solution of reference compound I-201 (1 equivalent) in tetrahydrofuran, ethanol and water (3:1:1 ratio) was treated with lithium hydroxide monohydrate (1.5 equivalents) and stirred at room temperature for 4 hours. at which time the solution was diluted with water and dichloromethane. The layers were separated and the aqueous layer was acidified to pH ~1. The acidified aqueous layer was extracted with dichloromethane, dried with magnesium sulfate, filtered, and the solvent was removed in vacuo to give the reference compound I-203 (18.5 mg, 65% yield) as a white solid.

1H-NMR (400 MHz, CD3OD) 88.76 (d, 1H), 8.15 (d, 1H), 7.36 (s, 1H), 7.29-7.24 (m, 1H), 7 .10-7.01 (m, 2H), 6.86-6.82 (m, 2H), 5.94 (s, 2H), 4.47 (s, 2H), 3.96 (d, 2H ), 1.22-1.15 (m, 1H), 0.59-0.55 (m, 2H), 0.38-0.34 (m, 2H).

Reference compound I-204

The title compound was prepared following general procedure B, 2-(isopropylamino)acetic acid was the amine reagent, and the reaction was heated at 100 °C for 16 h as a solution in dioxane/water (3:1). . The crude material was purified by silica gel chromatography (0-10% methanol in dichloromethane) to provide the desired compound, reference compound I-204 (8 mg, 33% yield) as a white solid.

1H-NMR (400 MHz, CDCla) 88.40 (br s, 1H), 8.11 (br s, 1H), 7.22 (s, 1H), 7.20-7.15 (m, 1H) , 7.20-6.92 (m, 2H), 6.83 (brs, 1H), 6.60 (brs, 1H), 5.91 (brs, 2H), 4.75 (brs, 1H), 4.14 (brs, 2H), 1.29 (d, 6H).

Reference compound I-205

The title compound was prepared following general procedure B, ethyl 2-(isobutylamino)acetate was the amine reagent, and the reaction was heated at 90 °C for 44 h as a solution in dioxane. After completion of the reaction, as determined by LC/MS, the solution was diluted with water and dichloromethane. The layers were separated and the aqueous layer was extracted with dichloromethane. The organic substances were dried over magnesium sulfate, filtered, and the solvent was removed in vacuo. The crude material was purified by silica gel chromatography (0-50% ethyl acetate in hexanes) to provide the desired compound, reference compound I-205 (19 mg, 57% yield) as a yellow solid.

1H-NMR (400 MHz, CDCh) 88.43 (m, 1H), 8.16 (d, 1H), 7.20-7.14 (m, 2H), 7.02-6.98 (m, 1H), 6.94 (t, 1H), 6.81 (t, 1H), 6.54 (m, 1H), 6.54 (m, 1H), 5.93 (s, 2H), 4, 26 (s, 2H), 4.16 (q, 2H), 3.54 (d, 2H), 2.09-2.03 (m, 1H), 1.19 (t, 3H), 0.97 (d, 6H).

Reference compound I-209

A solution of the reference compound I-205 (1 equivalent) in tetrahydrofuran, ethanol and water (3:1:1 ratio) was treated with lithium hydroxide monohydrate (2 equivalents) and stirred at room temperature until complete consumption of material. starting point, as determined by LC/MS. The solution was diluted with water and diethyl ether. The layers were separated and the aqueous layer was acidified to pH ~1. The acidified aqueous layer was extracted with dichloromethane, dried with ethyl acetate, filtered, and the solvent was removed in vacuo to give the reference compound I-209 (13 mg, 86% yield) as a white solid.

1H-NMR (400 MHz, CD3OD) 88.76 (m, 1H), 8.15 (d, 1H), 7.36 (s, 1H), 7.29-7.24 (m, 1H), 7 .10-7.01 (m, 2H), 6.86-6.82 (m, 2H), 5.92 (s, 2H), 4.38 (s, 2H), 3.61 (d, 2H ), 2.15-2.08 (m, 1H), 1.00 (d, 6H).

I-257 REFERENCE COMPOSITION

The title compound was prepared in 4 steps:

Stage 1: Synthesis of (S)-2-(4-methylphenylsulfonamido)-3-(pyridin-2-yl)propanoic acid

A suspension of (S)-2-amino-3-(pyridin-2-yl)propanoic acid, p-toluenesulfonic chloride (1.2 equiv.) in water was treated with sodium hydroxide (1 N solution, 3 equiv. ). The reaction was stirred at 90 °C for 18 h. The resulting mixture was cooled to room temperature, neutralized to pH 6 with 1 N HCl solution. Sodium chloride was added to saturate the solution which was then extracted with dichloromethane//so-propanol (4:1 v/v). . The combined organic phases were dried over sodium sulfate, filtered, and the solvent was removed in vacuo to provide (S)-2-(4-methylphenylsulfonamido)-3-(pyridin-2-yl)propanoic acid, an off-white solid (yield 35%).

Stage 2: Synthesis of (S)-2-(N,4-dimethylphenylsulfonamido)-3-(pyridin-2-yl)propanoic acid

A suspension of (S)-2-(4-methylphenylsulfonamido)-3-(pyridin-2-yl)propanoic acid and methyl iodide (3.2 equiv.) in aqueous 1 N sodium hydroxide solution (4.0 equiv.) was heated at 100 °C for 6 h. The resulting mixture was cooled to room temperature, neutralized to pH 6 with 1 N HCl solution and extracted with dichloromethane//so-propanol (4:1 v/v). The combined organic phases were dried over sodium sulfate, filtered, and the solvent was removed in vacuo. Purification by silica gel chromatography (gradient 20-50% acetonitrile/methanol (7:1) in dichloromethane) afforded (S)-2-(A/,4-dimethylphenylsulfonamido)-acid. 3-(pyríd¡n-2-¡l) propanoic as a yellow foam solid (yield 39%).

Step 3: Synthesis of (S)-2-(methylamino)-3-(pyridin-2-yl)propanoic acid (as HBr salt)

To (S)-2-(A/,4-dimethylphenylsulfonamido)-3-(pyridin-2-yl)propanoic acid was added HBr (33% by weight in glacial acetic acid, 25 equiv.). The reaction was stirred at 85 °C for 6 h and then at 60 °C for 3 d. An additional amount of HBr solution (3.3 equiv.) was added and the resulting mixture was stirred at 85 °C for 6 h. The reaction was cooled to room temperature, diluted with water and washed with ether. The aqueous phase was concentrated in vacuo providing (S)-2-(methylamino)-3-(pyridin-2-yl)propanoic acid dihydrobromide as a thick red oil (yield >99%) which was used in the next step without additional manipulation.

Stage 4: Synthesis of reference compound I-257

The title compound was prepared following general procedure B, except that (S)-2-(methylamino)-3-(pyridin-2-yl)propanoic acid (as the HBr salt) was the amine reagent, used 1.1 equivalents of intermediate 1, and the contents were heated to 100 °C. The crude material was purified by reverse phase HPLC (gradient of 20-75% acetonitrile in water with 0.1% trifluoacetic acid, 20 minutes) providing the desired compound, reference compound I-257 (123 mg, yield 73 %) as a tan solid.

1H-NMR (400 MHz,CDaOD) d 8.81 (d, 1H), 8.49 (d, 1H), 8.24 (d, 1H), 8.13 (ap. t, 1H), 7, 88 (d, 1H), 7.58 (ap. t, 1H), 7.42 (s, 1H), 7.29 (ap. q, 1H), 7.11 (m, 1H), 7.06 (ap . t, 1H), 6.90 (d, 1H), 6.89 (m, 1H), 6.01 (d, 1H), 5.97 (d, 1H), 5.65 (br. m, 1H), 3.88 (dd, 1H), 3.67 (dd, 1H), 3.35 (d, 3H).

1H-NMR (400 MHz, CD3OD) 88.81 (d, 1H), 8.49 (d, 1H), 8.24 (d, 1H), 8.13 (ap. t, 1H), 7.88 (d, 1H), 7.58 (ap. t, 1H), 7.42 (s, 1H), 7.29 (ap. q, 1H), 7.11 (m, 1H), 7.06 (ap. t, 1H), 6.90 (d, 1H), 6.89 (m, 1H), 6.01 (d, 1H), 5.97 (d, 1H), 5.65 (br. m, 1H ), 3.88 (dd, 1H), 3.67 (dd, 1H), 3.35 (d, 3H).

Reference compound I-200

The composition of the title was synthesized in 8 stages:

Step 1: Synthesis of ethyl 3-(soxazol-3-yl)-1H-1,2,4-triazole-5-carboxylate

A suspension of isoxazol-3-carbohydrazide (1 equiv.), ethyl 2-amino-2-thioxoacetate (1 equiv.) and ammonium chloride (10 equiv.) in absolute ethanol in a sealed vial was heated to 110 °C. for 7 d. The crude mixture was concentrated in vacuo. Water was added and the aqueous phase was extracted with ethyl acetate. The combined organic phases were dried over sodium sulfate, filtered, and the solvent was removed in vacuo. Purification by silica gel chromatography (10-20% acetonitrile/methanol (7:1) gradient in dichloromethane) afforded 3-(isoxazol-3-yl)-1H-1,2,4-tnazol-5-carboxylate. of ethyl as an orange solid (24% yield).

Step 2: Synthesis of ethyl 1-(2-fluorobenzyl)-5-(¡soxazol-3-¡l)-1H-1,2,4-t^azol-3-carboxylate and 1-( Ethyl 2-fluorobenzyl)-3-(¡soxazol-3-¡l)-1H-1,2,4-t^azol-5-carboxylate

To ethyl 3-(soxazol-3-l)-1H-1,2,4-t^azol-5-carboxylate in DMF was added sodium hydride (60% by weight in mineral oil, 1. 2 equiv.). After 10 min, 2-fluorobenzyl bromide (1.2 equiv.) was added and the reaction was stirred for 2 h. Water was added and the resulting mixture was extracted with ethyl acetate. The combined organic phases were washed with water and brine, dried over sodium sulfate, filtered, and the solvent was removed in vacuo. Purification by silica gel chromatography (10-40% ethyl acetate/hexanes gradient) afforded 1-(2-fluorobenzyl)-5-(¡soxazol-3-¡l)-1H-1,2, ethyl 4-t^azol-3-carboxylate and ethyl 1-(2-fluorobenzyl)-3-(isoxazol-3-yl)-1H-1,2,4-triazole-5-carboxylate (yield 63 %, ratio 42:58).

Stage 3: Synthesis of 1-(2-fluorobenzyl)-5-(¡soxazol-3-¡l)-1H-1,2,4-t^azol-3-carboxylic acid and 1-( 2-fluorobenzyl)-3-(isoxazol-3-yl)-1 H-1,2,4-triazole-5-carboxylic

To a solution of ethyl 1-(2-fluorobenzyl)-5-(¡soxazol-3-¡l)-1H-1,2,4-t^azol-3-carboxylate and 1-(2 Ethyl -fluorobenzyl)-3-(soxazol-3-l)-1H-1,2,4-triazole-5-carboxylate in tetrahydrofuran/methanol/water (3:1:1 ratio) is added hydrated lithium hydroxide (1.5 equivalents). After 1 h, water and 1 N HCl solution (50:8 ratio) were added and the resulting mixture was extracted with ethyl acetate. Note: The product was not completely soluble and was collected by vacuum filtration. The aqueous layer was extracted with dichloromethane//so-propanol (4:1 v/v). The combined organic phases were concentrated in vacuo and triturated with ether to provide additional product. The combined solids (88%, mixture of regioisomers) were used in the next step without further manipulation.

Step 4: Synthesis of 1-(2-fluorobenzyl)-5-(soxazol-3-l)-1H-1,2,4-triazole-3-carbonitrile

To a suspension of 1-(2-fluorobenzyl)-5-(isoxazol-3-¡l)-1H-1,2,4-triazole-3-carboxylic acid and 1-(2- fluorobenzyl)-3-(soxazol-3-yl)-1H-1,2,4-triazole-5-carboxylic acid, 2-methylpropan-2-amine (2 equiv.), and triethylamine (2 equiv. .) n-propylphosphonic anhydride (T3P, 50% solution in ethyl acetate, 3 equiv.) was added to ethyl acetate. The resulting yellow solution was heated at 65 °C for 2.5 h. The solvent was removed in vacuo. Phosphoryl trichloride (18 equiv.) was added and the resulting mixtures were stirred at 70 °C for 50 min. The reaction was quenched by carefully pouring into a mixture of ice and water, neutralized to pH 7 by the addition of saturated sodium bicarbonate solution, and extracted with dichloromethane. The combined organic phases were dried over sodium sulfate, filtered, and the solvent was removed in vacuo. Purification by silica gel chromatography (10-60% ethyl acetate/hexanes gradient) afforded 1-(2-fluorobenzyl)-5-(isoxazol-3-¡l)-1H-1,2,4-tr azol-3-carbonitrile (yield 39%). Note: One of the regioisomers was decarboxylated forming 3-(1-(2-fluorobenzyl)-1H-1,2,4-triazol-3-¡l)¡soxazole. The structural assignment is consistent with the observed nOe. This secondary reaction could have occurred during the saponification stage (stage 3).

Step 5: Synthesis of 1-(2-fluorobenzyl)-5-(soxazol-3-l)-1H-1,2,4-triazole-3-carboxymida

A solution of 1-(2-fluorobenzyl)-5-(¡soxazol-3-¡l)-1H-1,2,4-triazole-3-carbonitrile in methanol was treated with sodium methoxide (25 wt% solution in methanol, 5 equiv.) and stirred for 1 h. Ammonium chloride (10 equiv.) was added. After 18 h, the reaction mixture was concentrated in vacuo and partitioned between semisaturated sodium bicarbonate/1 N sodium hydroxide solution (10:1 ratio) and ethyl acetate. The organic phases were dried over sodium sulfate, filtered, and the solvent was removed in vacuo to provide 1-(2-fluorobenzyl)-5-(¡soxazol-3-¡l)-1H-1,2,4-triazole. -3-carboximidamide (yield >99%) which was used without further purification.

Step 6: Synthesis of 5-fluoro-2-(1-(2-fluorobenzyl)-5-(isoxazol-3-¡l)-1H-1,2,4-tr¡azol-3-¡l) pyrim¡ d¡n-4(3H)-one

A suspension of 1-(2-fluorobenzyl)-5-(¡soxazol-3-¡l)-1H-1,2,4-triazole-3-carboxymide was treated with (Z) -3-ethoxy¡-2-fluoro-3-oxoprop-1-en-1-olate (3.0 equiv.) and heated at 90 °C for 1 h. After cooling to room temperature, the reaction mixture was neutralized by the addition of HCl (1.25 M solution in ethanol). The resulting pale yellow suspension was stirred for 5 min and then concentrated in vacuo. The residue was partitioned between dichloromethane and water and the aqueous layer was extracted again with dichloromethane. The combined organic phases were dried over sodium sulfate, filtered, and the solvent was removed in vacuo. Purification by silica gel chromatography (gradient of 5-20% acetonitrile/methanol (7:1) in dichloromethane) afforded 5-fluoro-2-(1-(2-fluorobenzyl)-5-(¡soxazol-3- ¡l)-1H-1,2,4-tr¡azol-3-¡l)-pyr¡m¡d¡n-4(3H)-one (61 mg, 74% yield) as a yellow solid pale.

1H-NMR (400 MHz, CD3OD) 58.95 (d, 1H), 8.09 (d, 1H), 7.35 (ap. q, 1H), 7.23 (ap. t, 1H), 7 .17 (d, 1H), 7.15 (m, 2H), 6.07 (s, 2H).

Step 7: Synthesis of 3-(3-(4-chloro-5-fluoropyrimidin-2-yl)-1-(2-fluorobenzyl)-1H-1,2,4-triazol-5-yl)isoxazole

A suspension of 5-fluoro-2-(1-(2-fluorobenzyl)-5-(isoxazol-3-yl)-1H-1,2,4-triazol-3-yl)pyrimidin-4(3H)-one in phosphoryl trichloride (77 equiv.) was heated at 65 °C for 2 h. The reaction mixture was carefully poured onto ice and stirred for 20 min. The resulting mixture was made basic to pH 8 by the addition of saturated sodium bicarbonate and extracted with dichloromethane. The combined organic layers were dried over sodium sulfate, filtered, and the solvent was removed in vacuo to provide 3-(3-(4-chloro-5-fluoropyrimidin-2-yl)-1-(2-fluorobenzyl)-1H -1,2,4-triazol-5-yl)isoxazole as an off-white solid (yield >99%).

Step 8: Synthesis of reference compound I-200

The title compound was prepared following general procedure B, except that (S)-3-methyl-2-(methylamino)butanoic acid was the amine reagent, 3-(3-(4-chloro-5-) was used fluoropyrimidin-2-yl)-1-(2-fluorobenzyl)-1H-1,2,4-triazol-5-yl)isoxazole instead of intermediate 1, and the contents were heated to 100 °C. The crude material was purified by silica gel chromatography (20-50% gradient (acetonitrile/methanol=7:1) in dichloromethane) obtaining an impure product. The sample was purified again by reverse phase HPLC (gradient of 95% acetonitrile in water with 0.1% trifluoacetic acid, 20 minutes) to provide the desired compound, reference compound I-200 (23 mg, 66% in two stages) as a white solid.

1H-NMR (400 MHz, CDCla) 58.56 (d, 1 H), 8.34 (d, 1 H), 7.27 (ap. q, 1 H), 7.21 (ap. t, 1 H), 7.15 (d, 1 H), 7.06 (m, 2 H), 6.09 (d, 1 H), 6.02 (d, 1 H), 4.27 (d, 1 H), 3.07 (d, 3 H), 2.55 (m, 1 H), 1.10 (d, 3 H), 0.95 (d, 3 H).

Reference compound I-249

The title compound was prepared following general procedure B, except that 4-methylpiperidine-4-carboxylic acid (as the HCl salt) was the amine reagent, 3-(3-(4-chloro-5- fluoropyrimidin-2-yl)-1-(2-fluorobenzyl)-1H-1,2,4-triazol-5-yl)isoxazole (the synthesis of which is described in the procedure described for the reference compound I-200) instead of intermediate product 1 and the content was heated at 100 °C for 19 h. The reaction was cooled and diluted with water, and neutralized with aqueous 1N HCl. The resulting solids were collected by vacuum filtration, washed with water, and dried under vacuum to provide the desired compound, reference compound I-249 (29 mg, 85% yield) as an off-white solid.

1H-NMR (400 MHz, CDCl3) 58.54 (d, 1H), 8.22 (d, 1H), 7.24 (m, 1H), 7.17 (d, 1H), 7.07-7 .00 (m, 3H), 6.08 (s, 2H), 4.41 (br. d, 2H), 3.38 (ap. t, 2H), 2.28 (br. d, 2H), 1.59 (m, 2H), 1.31 (s, 3H).

Reference compound I-1

The title compound was prepared following general procedure B, except that pyrrolidine (7 equiv.) was the amine reagent, triethylamine was not used, and the contents were heated at 40 °C for 10 min as a solution in THF, then at 23 °C for 18 h. The reaction was cooled and diluted with ethyl acetate, and washed with water and brine. The solvent was removed in vacuo providing the desired compound, reference compound I-1 (23 mg, 76% yield) as a solid.

1H NMR (400 MHz, DMSO-d6) 5 ppm 9.08 (d, 1 H), 8.20 (d, 1 H), 7.52 (s, 1 H), 7.30 - 7.40 ( m, 1 H), 7.18 - 7.28 (m, 2 H), 7.10 (t, 1 H), 6.82 (t, 1 H), 5.89 (s, 2 H), 3.65 - 3.70 (m, 4 H), 1.92 (d, 4 H).

Reference compound I-2

The title compound was prepared following general procedure B, except that piperidine (7 equiv.) was the amine reagent, triethylamine was not used, and the contents were heated at 40 °C for 10 min as a solution in THF, then at 23 °C for 18 h. The reaction was cooled and diluted with ethyl acetate, and washed with water and brine. The solvent was removed in vacuo providing the desired compound, reference compound I-2 (25 mg, 80% yield) as a solid.

1H NMR (400 MHz, DMSO-da) 5 ppm 9.09 (s, 1 H), 8.26 (d, 1 H), 7.54 (s, 1 H), 7.32 (s, 2 H) ), 7.20 - 7.27 (m, 2 H), 7.10 (t, 1 H), 6.81 (t, 1 H), 5.90 (s, 2 H), 3.74 - 3.80 (m, 4 H), 1.58 - 1.62 (m, 6 H).

Reference compound I-3

The title compound was prepared following general procedure B, except that morpholine (7 equiv.) was the amine reagent, no triethylamine was used, and the contents were heated at 40 °C for 10 min as a solution in THF, then at 23 °C for 18 h. The reaction was cooled and diluted with ethyl acetate, and washed with water and brine. The solvent was removed in vacuo providing the desired compound, reference compound I-3 (24 mg, 76% yield) as a solid.

1H NMR (400 MHz, DMSO-d6) 5 ppm 9.08 (s, 1 H), 8.33 (d, 1 H), 7.57 (s, 1 H), 7.32 (d, 2 H ), 7.20 - 7.27 (m, 2 H), 7.10 (t, 2 H), 6.81 (t, 1 H), 5.90 (s, 2 H), 3.79 ( d, 4 H), 3.74 (d, 4 H).

Reference compound I-4

The title compound was prepared following general procedure B, except that piperazine (7 equiv.) was the amine reagent, triethylamine was not used, and the contents were heated at 40 °C for 10 min as a solution in THF, then at 23 °C for 18 h. The reaction was cooled and diluted with ethyl acetate, and washed with water and brine. The solvent was removed in vacuo providing the desired compound, reference compound I-4 (20 mg, 64% yield) as a solid.

1H NMR (400 MHz, DMSO-d6) 5 ppm 9.09 (s, 1 H), 8.33 (d, 1 H), 7.56 (s, 1 H), 7.32 (s, 1 H) ), 7.20 - 7.28 (m, 2 H), 7.10 (t, 1 H), 6.81 (t, 1 H), 5.90 (s, 2 H), 3.78 - 3.84 (m, 4 H), 2.90 - 3.00 (m, 3 H).

Reference compound I-5

The title compound was prepared following general procedure B, except that N-methylpiperazine (7 equiv.) was the amine reagent, triethylamine was not used, and the contents were heated to 40 °C for 10 min as a solution in THF, then at 23 °C for 18 h. The reaction was cooled and diluted with ethyl acetate, and washed with water and brine. The solvent was removed in vacuo providing the desired compound, reference compound I-5 (23 mg, 71% yield) as a solid.

1H NMR (400 MHz, DMSO-d6) 5 ppm 9.09 (s, 1 H), 8.31 (d, 1 H), 7.56 (s, 1 H), 7.32 (s, 1 H) ), 7.18 - 7.27 (m, 2 H), 7.10 (t, 1 H), 6.80 (s, 1 H), 5.90 (s, 2 H), 3.74 - 3.81 (m, 4 H), 3.25 - 3.35 (s, 3 H), 2.20 - 2.30 (m, 4 H).

Reference compound I-6

The title compound was prepared following general procedure B, except that 2-morpholinoethanamine (7 equiv.) was the amine reagent, triethylamine was not used, and the contents were heated at 40 °C for 10 min as a solution in THF, then at 23 °C for 18 h. The reaction was cooled and diluted with ethyl acetate, and washed with water and brine. The solvent was removed in vacuo providing the desired compound, reference compound I-6 (25 mg, 72% yield) as a gum.

1H NMR (400 MHz, DMSO-d6) 5 ppm 9.09 (d, 1 H), 8.17 (d, 1 H), 7.59 (s, 1 H), 7.48 (s, 1 H) ), 7.28 - 7.38 (m, 1 H), 7.17 - 7.26 (m, 2 H), 7.10 (t, 1 H), 6.86 (t, 1 H), 5.88 (s, 2 H), 3.56 - 3.62 (m, 4 H), 3.48 (t, 4 H), 2.44 (m, 2 H), 2.29 - 2, 40 (m, 2 H).

Reference compound I-7

The title compound was prepared following general procedure B, except that W,A/-dimethylethane-1,2 diamine (7 equiv.) was the amine reagent, no triethylamine was used, and the contents were heated to 40° C for 10 min as a solution in THF, then at 23 °C for 18 h. The reaction was cooled and diluted with ethyl acetate, and washed with water and brine. The solvent was removed in vacuo providing the desired compound, reference compound I-7 (24 mg, 76% yield) as a gum.

1H NMR (400 MHz, DMSO-d6) 5 ppm 9.09 (d, 1 H), 8.17 (d, 1 H), 7.54 (s, 1 H), 7.48 (s, 1 H) ), 7.29 - 7.38 (m, 1 H), 7.18 - 7.27 (m, 2 H), 7.11 (t, 1 H), 6.88 (t, 1 H), 5.89 (s, 2 H), 3.57 (q, 2 H), 2.43-2.49 (m, 2 H), 2.19 (s, 6 H).

Reference compound I-9

The title compound was prepared following general procedure B, except that cyclohexylamine (7 equiv.) was the amine reagent, triethylamine was not used, and the contents were heated to 40 °C for 10 min as a solution in THF, then at 23 °C for 18 h. The reaction was cooled and diluted with ethyl acetate, and washed with water and brine. The solvent was removed in vacuo providing the desired compound, reference compound I-9 (20 mg, 62% yield) as a solid.

1H NMR (400 MHz, DMSO-da) 5 ppm 9.09 (d, 1 H), 8.14 (d, 1 H), 7.51 (d, 1 H), 7.45 (s, 1 H) ), 7.33 (d, 1H), 7.18 - 7.28

(m, 2 H), 7.10 (t, 1 H), 6.85 (t, 1 H), 5.88 (s, 2 H), 4.03-4.08 (m, 1 H) , 1.89-1.92 (m, 2 H), 1.72-1.76 (m, 2 H), 1.63 (d, 2

H), 1.32- 1.43 (m, 4 H).

Reference compound I-8

The title compound was prepared following general procedure B, except that dimethylamine (7 equiv.) was the amine reagent, triethylamine was not used, and the contents were heated to 40 °C for 10 min as a solution in THF, then at 23 °C for 18 h. The reaction was cooled and diluted with ethyl acetate, and washed with water and brine. The solvent was removed in vacuo providing the desired compound, reference compound I-8 (19 mg, 67% yield) as a solid.

1H NMR (400 MHz, DMSO-d6) 5 ppm 9.09 (s, 1 H), 8.18 (d, 1 H), 7.52 (s, 1 H), 7.39 - 7.45 ( m, 1 H), 7.18 - 7.27 (m, 2

H), 7.10 (t, 1 H), 6.82 - 6.88 (m, 1 H), 5.90 (s, 2 H), 3.24 (d, 6 H).

Reference compound I-11

The title compound was prepared following general procedure B, except that 2-methylpyrrolidine was the amine reagent, triethylamine was not used, and the contents were heated at 40 °C for 10 min as a solution in THF, then 23 °C for 18 h. The reaction was cooled and diluted with ethyl acetate, and washed with water and brine.

The solvent was removed in vacuo, and the crude material was purified by silica gel chromatography using a 0-10% methanol/dichloromethane gradient providing the desired compound, reference compound I-11.

(16 mg, 51% yield) as a solid.

1H NMR (400 MHz, DMSO-d6) 5 ppm 9.09 (d, 1 H), 8.21 (d, 1 H), 7.49 (s, 1 H), 7.29 - 7.38 ( m, 1 H), 7.17 - 7.27 (m, 2

H), 7.10 (t, 1 H), 6.84 (t 1 H), 5.92 (s, 2 H), 4.40-4.48 (m, 1 H), 3.75 - 3.90 (m, 1 H), 3.56 - 3.69 (m, 1 H), 2.00 - 2.07

(m, 2 H), 1.93 (d, 1 H), 1.65-1.73 (m, 1 H), 1.23 (d, 3 H).

Reference compound I-10

The title compound was prepared following general procedure B, except that piperidin-4-ol was the amine reagent, no triethylamine was used, and the contents were heated at 40 °C for 10 min as a solution in THF, at then at 23 °C for 18 h. The reaction was cooled and diluted with ethyl acetate, and washed with water and brine.

The solvent was removed in vacuo providing the desired compound, reference compound I-10 (19 mg, 58% yield) as a solid.

1H NMR (400 MHz, DMSO-d6) 5 ppm 9.08 (d, 1 H), 8.27 (d, 1 H), 7.54 (s, 1 H), 7.21 - 7.39 ( m, 3 H), 7.10 (t, 1 H), 6.81

(t, 1 H), 5.90 (s, 2 H), 4.78-4.84 (m, 1 H), 4.18 (d, 2 H), 3.74-3.79 (m , 1 H), 3.37 - 3.47 (m, 2 H), 1.81-8.89 (m, 2 H),

1.40 - 1.54 (m, 2H).

Reference compound I-12

The title compound was prepared following general procedure B, except that fed-butyl 4-aminopiperidine-1-carboxylate (1.5 equiv.) was the amine reagent, no triethylamine was used, and the contents were heated to 40 °C for 10 min as a solution in THF, then at 23 °C for 18 h. The reaction was cooled and diluted with ethyl acetate, and washed with water and brine. The solvent was removed in vacuo, and the crude material was purified by silica gel chromatography using a gradient of 0-10% methanol/dichloromethane providing the desired compound, reference compound I-12 (36 mg, 90% yield). like a solid.

1H NMR (400 MHz, DMSO-d6) 5 ppm 9.04 - 9.15 (m, 1 H), 8.18 (d, 1 H), 7.53 - 7.65 (m, 1 H), 7.47 (s, 1 H), 7.30 - 7.39

(m, 1 H), 7.19 - 7.27 (m, 2 H), 7.07 - 7.16 (m, 1 H), 6.85 (t, 1 H), 5.83 - 5 .91 (m, 2 H), 4.25 (d, 1 H), 3.96 (d, 2 H), 2.87 - 2.91 (m, 2 H), 1.87 (d, 2 H), 1.44 -1.51 (m, 2 H),1.41 (s, 9 H).

Reference compound I-13

The title compound was prepared following general procedure B, except that (S)-pyrrolidin-2-ylmethanol was the amine reagent, no triethylamine was used, and the contents were heated to 40 °C for 10 min as a solution. in

THF, then at 23 °C for 18 h. The reaction was cooled and diluted with ethyl acetate, and washed with water and brine. The solvent was removed in vacuo, and the crude material was purified by silica gel chromatography using a 0-10% methanol/dichloromethane gradient to provide the desired compound, reference compound I-13 (18 mg, 55% yield). like a solid.

1H NMR (400 MHz, DMSO-d6) 5 ppm 9.09 (d, 1 H), 8.21 (d, 1 H), 7.49 (s, 1 H), 7.28 - 7.37 ( m, 1 H), 7.18 - 7.26 (m, 2

H), 7.10 (t, 1 H), 6.83 (t, 1 H), 5.85 - 5.93 (m, 2 H), 4.86 (t, 1 H), 4.32 - 4.39 (m, 1 H), 3.74 - 3.79 (m, 1 H), 3, (m, 1 H), 3.52 - 3.59 (m, 1 H), 3, 44 - 3.50 (m, 1 H), 1.98 - 2.04 (m, 2 H), 1.91 (d, 2 H).

REFERENCE COMPOSITION I-17

The title compound was prepared following general procedure B, except that 3-methoxypyrrolidine (4 equiv.) was the amine reagent, and 4 equivalents of triethylamine were used, and the contents were heated to 40 °C for 1 h as a solution in THF. The reaction was cooled and diluted with ethyl acetate, and washed with water and brine. The solvent was removed in vacuo providing the desired compound, reference compound I-17 (12 mg, 61% yield) as a solid.

1H NMR (400 MHz, DMSO-da) 8 ppm 9.09 (d, 1 H), 8.23 (d, 1 H), 7.53 (s, 1 H), 7.29 - 7.40 ( m, 1 H), 7.18 - 7.28 (m, 2 H), 7.10 (t, 1 H), 6.82 (t, 1 H), 5.90 (s, 2 H), 4.03 - 4.12 (m, 1 H), 3.70 - 3.87 (m, 3 H), 3.66 (d, 1 H), 3.28 (s, 3 H), 1, 96 - 2.15 (m, 2H).

Reference compound I-18

The title compound was prepared following general procedure B, except that piperidin-3-ol (4 equiv.) was the amine reagent, no triethylamine was used, and the contents were heated to 40 °C for 1 h as a solution in THF. The reaction was cooled and diluted with ethyl acetate, and washed with water and brine. The solvent was removed in vacuo providing the desired compound, reference compound I-18 (14 mg, 72% yield) as a solid.

1H NMR (400 MHz, DMSO-d6) 8 ppm 9.09 (d, 1 H), 8.25 (d, 1 H), 7.53 (s, 1 H), 7.28 - 7.39 ( m, 1 H), 7.17 - 7.27 (m, 2 H), 7.10 (t, 1 H), 6.81 (t, 1 H), 5.90 (s, 2 H), 4.97 (d, 1 H), 4.18 (d, 1 H), 3.56 - 3.68 (m, 1 H), 3.37 - 3.48 (m, 2 H), 3, 21 (dd, 1 H), 1.73 - 1.96 (m, 2 H), 1.43-1.58 (m, 2 H).

Reference compound I-25

The title compound was prepared following general procedure B, except that detert-butyl piperidin-3-ylcarbamate (4 equiv.) was the amine reagent, no triethylamine was used, and the contents were heated to 40 °C for 1 h as a solution in THF. The reaction was cooled and diluted with ethyl acetate, and washed with water and brine. The solvent was removed in vacuo, and the crude material was purified by silica gel chromatography using a gradient of 0-10% methanol/dichloromethane providing the desired compound, reference compound I-25 (25 mg, 66% yield) as a solid.

1H NMR (500 MHz, CD3OD) 8 ppm 8.75 (d, 1 H), 8.16 (d, 1 H), 7.58 (s, 1 H), 7.23 - 7.38 (m, 1 H), 7.08 - 7.17 (m, 1 H), 7.05 (t, 1 H), 6.95 (s, 1 H), 6.84 (t, 1 H), 6.78 ( d, 1 H), 5.98 (s, 2 H), 4.40 (d, 1 H), 4.13 - 4.24 (m, 1 H), 3.69 (br. s., 1 H), 3.56 (d, 1 H), 3.35 - 3.42 (m, 1 H), 2.00 - 2.09 (m, 1 H), 1.91 (dd, 1 H) , 1.59 - 1.72 (m, 2 H), 1.45 (s, 9 H).

Reference compound I-26

The title compound was prepared following general procedure B, except that tert-butyl 3-aminoazetidine-1-carboxylate (4 equiv.) was the amine reagent, 2 equivalents of triethylamine were used, and the contents were heated to 40 °C for 1 h as a solution in THF, followed by heating at 75 °C until complete consumption of starting material was observed by LC/MS. The reaction was cooled and diluted with ethyl acetate, and washed with water and brine. The solvent was removed in vacuo, and the crude material was purified by silica gel chromatography using a 0-10% methanol/dichloromethane gradient to provide the desired compound, reference compound I-26 (24 mg, 67% yield). like a solid.

1H NMR (400 MHz, DMSO-d6) 8 ppm 9.10 (d, 1 H), 8.31 (d, 1 H), 8.25 (d, 1 H), 7.48 - 7.57 ( m, 1 H), 7.29 - 7.38 (m, 1 H), 7 .15-7.28 (m, 2 H), 7.11 (t, 1 H), 6.85 (t, 1 H), 5.89 (s, 2 H), 4.76-4.93 (m, 1 H), 4.15-4.25 (m, 2 H), 3.91 (dd, 2 H ), 1.39 (s, 9H).

Reference compound I-27

The title compound was prepared following general procedure B, except that tert-butyl 3-aminopiperidine-1-carboxylate (4 equiv.) was the amine reagent, 2 equivalents of triethylamine were used, and the contents were heated to 40 °C for 1 h as a solution in THF, followed by heating at 75 °C until complete consumption of starting material was observed by LC/MS. The reaction was cooled and diluted with ethyl acetate, and washed with water and brine. The solvent was removed in vacuo, and the crude material was purified by silica gel chromatography using a 0-10% methanol/dichloromethane gradient to provide the desired compound, reference compound I-27 (24 mg, 57% yield). like a solid.

1H NMR (500 MHz, CD3OD) 8 ppm 8.78 (d, 1 H), 8.15 (d, 1 H), 7.58 (s, 1 H), 7.25 - 7.33 (m, 1 H), 7.08 - 7.17 (m, 1 H), 7.05 (t, 1 H), 6.95 (s, 1 H), 6.84 (t, 1 H), 6.78 ( d, 1 H), 5.98 (s, 2 H), 4.40 (d, 1 H), 4.18 (d, 1 H), 3.69 (m, 1 H), 3.56 ( d, 1 H), 3.39 (d, 1 H), 1.99-2.08 (m, 1H), 1.91 (dd, 1 H), 1.57- 1.76 (m, 2 H), 1.45 (s, 9 H).

Reference compound I-19

The title compound was prepared following general procedure B, except that 3-methoxypiperidine (4 equiv.) was the amine reagent, and the content was heated to 40 °C for 1 h as a solution in THF, followed by heating at 75 °C until complete consumption of starting material was observed in LC/MS. The reaction was cooled and filtered, and the solids were washed with ethyl acetate. The filtrate was collected and washed with water and brine. The solvent was removed in vacuo providing the desired compound, reference compound I-19 (15 mg, 74% yield) as a solid.

1H NMR (400 MHz, DMSO-da) 8 ppm 9.06 (d, 1 H), 8.23 (d, 1 H), 7.49 (s, 1 H), 7.25 - 7.35 ( m, 1 H), 7.13 - 7.24 (m, 2 H), 7.07 (t, 1 H), 6.78 - 6.87 (m, 1 H), 5.86 (s, 2 H), 3.91 (d, 1 H), 3.62 - 3.81 (m, 3 H), 3.34 (dd, 1 H), 3.23 (s, 3 H), 1, 83 - 1.94 (m, 1 H), 1.72 - 1.81 (m, 1 H), 1.56 - 1.65 (m, 1 H), 1.44 - 1.54 (m, 1 HOUR).

Reference compound I-20

The title compound was prepared following general procedure B, except that pyrrolidin-3-ol (4 equiv.) was the amine reagent, no triethylamine was used, and the contents were heated to 40 °C for 1 h as a solution in THF. The reaction was cooled and filtered, and the solids were washed with ethyl acetate. The filtrate was collected and washed with water and brine. The solvent was removed in vacuo providing the desired compound, reference compound I-20 (10 mg, 53% yield) as a solid.

1H NMR (400 MHz, DMSO-d6) 8 ppm 9.08 (d, 1 H), 8.21 (d, 1 H), 7.52 (s, 1 H), 7.29 - 7.37 ( m, 1 H), 7.18 - 7.28 (m, 2 H), 7.10 (t, 1 H), 6.82 (t, 1 H), 5.90 (s, 2 H), 5.04 (d, 1 H), 4.32-4.39 (m, 1 H), 3.81 (d, 1 H), 3.69-3.77 (m, 1 H), 3, 60 -3.68 (m, 1 H), 1.96-2.05 (m, 1 H), 1.88 - 1.95 (m, 1 H).

Reference compound I-21

The title compound was prepared following general procedure B, except that detert-butyl acetidin-3-ylcarbamate (4 equiv.) was the amine reagent, no triethylamine was used, and the contents were heated to 40 °C for 1 h as a solution in THF. The reaction was cooled and filtered, and the solids were collected and dried under vacuum to provide the desired compound, reference compound I-21 (30 mg, 79% yield) as a solid.

1H NMR (400 MHz, DMSO-d6) 8 ppm 9.08 (s, 1 H), 8.25 (m, 1 H), 7.62 - 7.66 (m, 2 H), 7.51 ( s, 1 H), 7.29 - 7.34 (m, 1 H), 7.15 - 7.27 (m, 2 H), 7.07 - 7.14 (m, 1 H), 6, 79 - 6.84 (m, 1 H), 5.90 (s, 2 H), 4.35 - 4.44 (m, 2 H), 4.01 - 4.12 (m, 2 H), 3.84-3.90 (m, 1 H), 1.39 (s, 9 H).

Reference compound I-22

The title compound was prepared following general procedure B, except that 4-methoxypiperidine (4 equiv.) was the amine reagent, no triethylamine was used, and the contents were heated to 40 °C for 1 h as a solution in THF. The reaction was cooled and diluted with ethyl acetate, and washed with water and brine. The solvent was removed in vacuo providing the desired compound, reference compound I-22 (15 mg, 74% yield) as a solid.

1H NMR (400 MHz, DMSO-d6) 8 ppm 9.05 (d, 1 H), 8.25 (d, 1 H), 7.51 (s, 1 H), 7.26 - 7.31 ( m, 1 H), 7.14 - 7.25 (m, 2 H), 7.07 (t, 1 H), 6.74 - 6.82 (m, 1 H), 5.87 (s, 2 H), 4.06 - 4.11 (m, 2 H), 3.42 - 3.50 (m, 2 H), 3.25 (s, 3 H), 1.89-1.95 ( m, 2 H), 1.46-1.51 (m, 3 H).

Reference compound I-64

The title compound was prepared following general procedure B, except that 4-methyltetrahydro-2H-pyran-4-amine (as the HCl salt) (5 equiv.) was the amine reagent, 10 equivalents of base were used of Hunig instead of triethylamine and the content was heated at 175 °C for 1 h in the microwave as a solution in THF/DMF (1:1). The reaction was cooled and diluted with ethyl acetate, and washed with water and brine. The solvent was removed in vacuo, and the crude material was purified by silica gel chromatography using a gradient of 5-90% ethyl acetate/hexanes providing the desired compound, reference compound I-64 (4 mg, yield 20 %) as a solid.

1H NMR (500 MHz, CD3OD) 8 ppm 8.74 (s, 1 H), 8.05 (d, 1 H), 7.19 - 7.33 (m, 2 H), 6.97 - 7, 12 (m, 2 H), 6.79 - 6.93 (m, 2 H), 5.92 (s, 2 H), 3.67 - 3.81 (m, 4 H), 2.49 ( d, 2 H), 1.77 -1.91 (m, 2 H), 1.63 (s, 3 H).

Reference compound I-101

The title compound was prepared following general procedure B, except that 4-(tert-butoxycarbonyl)piperazin-2-carboxylic acid (4 equiv.) was the amine reagent, 5 equivalents of triethylamine were used, and the contents were heated to 90 °C for 5 h as a solution in THF/water (9:1). The reaction was taken up with ethyl acetate instead of dichloromethane. The crude material was purified by reverse phase HPLC providing the desired compound, reference compound I-101 (12 mg, 26% yield) as a solid.

1H NMR (500 MHz, DMSO-d6) 8 ppm 9.10 (d, 1 H), 8.39 (d, 1 H), 7.54 (s, 1H), 7.28 - 7.39 (m , 1 H), 7.15 - 7.25 (m, 2 H), 7.11 (t, 1 H), 6.85 (t, 1 H), 5.85- 5.97 (m, 2 H), 5.17 (br. s., 1 H), 4.45 (d, 1 H), 4.33 (br. s., 1 H), 3.91 -4.02 (m, 1 H), 3.33 - 3.39 (m, 2 H), 3.05 - 3.17 (m, 1 H), 1.41 (s, 9 H).

REFERENCE COMPOSITION I-163

The title compound was prepared following general procedure B, except that 4,5,6,7-tetrahydro-3H-imidazo[4,5-c]pyridine-4-carboxylic acid (4 equiv.) was the reagent. amine, 3 equivalents of triethylamine were used, and the content was heated at 175 °C for 10 min as a solution in NMP. The contents were diluted with diethyl ether, and the resulting precipitate was filtered and collected. The crude material was further purified by reverse phase HPLC providing the desired compound, reference compound I-163 (5 mg, 15% yield) as a solid.

1H NMR (500 MHz, methanol-d^ 8 ppm 9.03 (d, 1 H), 8.82 - 8.89 (m, 1 H), 8.58 (s, 1 H), 7.58 ( s, 1 H), 7.28 - 7.37 (m, 1

H), 7.11 -7.18 (m, 1 H), 7 .02-7.10 (m, 1 H), 6.94 (s, 1 H), 6.83-6.92 (m , 1 H), 6.01 (s, 2 H), 5.12 (d, 1 H), 5.00 (d, 1

H), 4.55 - 4.64 (m, 1 H), 3.43 (dd, 1 H), 3.20 (d, 1 H).

Reference compound I-189

The title compound was prepared following general procedure B, except that 4-ethylpiperidine-4-carboxylic acid (as the HCl salt) was the amine reagent, 4 equivalents of triethylamine were used, and the contents were heated to 40

°C for 18 h as a solution in THF. The reaction was incomplete, so additional amine reagent (3 equiv.), triethylamine (4 equiv.), and DMF (equivalent volume in THF) was introduced into the vessel, and the resulting mixture was heated to 85 °C for 18 h. The reaction was cooled and diluted with ethyl acetate, and washed with water and brine.

The solvent was removed in vacuo and the crude material was purified by reverse phase HPLC to provide the desired compound, reference compound I-189 as a solid.

1H NMR (500 MHz, methanol-d^ 8 ppm 8.79 (d, 1 H), 8.20 (d, 1 H), 7.54 (s, 1 H), 7.21 - 7.35 ( m, 1 H), 7.07 - 7.13 (m, 1

H), 7.05 (t, 1 H), 6.95 (d, 1 H), 6.89 (t, 1 H), 5.99 (s, 2 H), 4.65 (d, 2 H), 3.33 - 3.43 (m, 2 H), 2.32 (d, 2 H), 1.63 - 1.68 (m, 2 H), 1.55 - 1.63 (m , 2H), 0.91 (t, 3H).

Reference compound I-190

The title compound was prepared following general procedure B, except that 3-methylpiperidine-4-carboxylic acid was the amine reagent, 4 equivalents of triethylamine were used, and the contents were heated to 40 °C for

18 h as a solution in THF. The reaction was incomplete, so additional amine reagent (3 equiv.), triethylamine (4 equiv.), and DMF (equivalent volume in THF) was introduced into the vessel, and the resulting mixture was heated to 85 °C for 18 h. The reaction was cooled and diluted with ethyl acetate, and washed with water and brine. The solvent was removed in vacuo and the crude material was purified by reverse phase HPLC to provide the desired compound, reference compound I-190 as a solid.

1H NMR (500 MHz, methane l^) 8 ppm 8.79 (d, 1 H), 8.22 - 8.29 (m, 1 H), 7.59 (s, 1 H), 7.24 -7.34 (m, 1 H), 7.02 -7.15 (m, 2 H), 6.96 - 7.00 (m, 1 H), 6.93 (t, 1 H), 6 .01 (m, 2 H), 4.76 (d, 1 H), 4.58 (d, 1 H), 3.70 (dd, 1 H), 3.52 - 3.58 (m, 1

H), 2.87-2.94 (m, 1 H), 2.45-2.54 (m, 1 H), 2.00-2.11 (m, 1 H), 1.90- 1 .99 (m, 1 H), 1.02 (d, 3 H).

Reference compound I-235

The title compound was prepared in 3 steps:

Step 1: Synthesis of 2-(1-(2,3-difluorobenzyl)-5-(isoxazol-3-yl)-1H-pyrazol-3-yl)-5-fluoropyrimidin-4(3H)-one

The above compound was prepared following general procedure A, using 1-(isoxazol-3-yl)ethanone in step 1 and 2,3-difluorobenzylhydrazine in step 2.

Step 2: Preparation of 3-(3-(4-chloro-5-fluoropyrimidin-2-yl)-1-(2,3-difluorobenzyl)-1H-pyrazol-5-yl)isoxazole

A suspension of 2-(1-(2,3-difluorobenzyl)-5-(isoxazol-3-yl)-1H-pyrazol-3-yl)-5-fluoropyrimidin-4(3H)-one in phosphoryl trichloride ( 47 equiv) was heated at 65 °C for 2 hours. The reaction mixture was carefully poured onto ice and stirred for 20 min. The resulting mixture was made basic to pH 8 by the addition of saturated sodium bicarbonate and extracted with dichloromethane. The combined organic layers were dried over sodium sulfate, filtered, and the solvent was removed in vacuo to provide 3-(3-(4-chloro-5-fluoropyrimidin-2-yl)-1-(2,3-difluorobenzyl) -1H-pyrazol-5-yl)isoxazole as a light yellow solid which was used in the next step without further manipulation.

Stage 3: Synthesis of reference compound I-235

A solution of (S)-3-methyl-2-(methylamino)butanoic acid (3.0 equivalents), triethylamine (10 equivalents) and 3-(3-(4-chloro-5-fluoropyrimidin-2-yl)- 1-(2,3-difluorobenzyl) 1H-pyrazol-5-yl)isoxazole was stirred in dioxane/water (2:1 ratio) at 100

°C for 23 h, following general procedure B. The solution was diluted with water, neutralized to pH 3 by the addition of 1N HCl and extracted with dichloromethane. The combined organic layers were dried over sodium sulfate, filtered, and the solvent was removed in vacuo. Purification by silica gel chromatography (gradient of 0

10% acetonitrile/methanol (7:1) in dichloromethane) gave the reference compound I-235 (38 mg, 61% over 2 steps) as an off-white solid.

1H-NMR (400 MHz, CDCla) 88.46 (d, 1H), 8.22 (d, 1H), 7.23 (s, 1H), 7.04 (dd, 1H), 6.92 (dd 1H), 6.77 (ap. t, 1H), 6.58

(d, 1H), 5.99 (d, 1H), 5.94 (d, 1H), 4.27 (d, 1H), 3.24 (d, 3H), 2.52 (m, 1H) , 1.11 (d, 3H), 0.94 (d, 3H).

Reference compound I-236

The title compound was prepared in 3 steps:

Step 1: Synthesis of 5-fluoro-2-(1-(2-fluorobenzyl)-5-(oxazol-2-N)-1H-pyrazole-3-N) pyrimidin-4(3H)-one

The above compound was prepared following general procedure A, using 1-(oxazol-2-yl)ethanone in step 1 and 2-fluorobenzylhydrazine in step 2.

Step 2: Synthesis of 2-(3-(4-chloro-5-fluoropyrimidin-2-yl)-1-(2-fluorobenzyl)-1H-pyrazol-5-yl)oxazole

The previous compound was prepared following a process analogous to step 2 of the synthesis of the reference compound I-235, using 5-fluoro-2-(1-(2-fluorobenzyl)-5-(oxazol-2-yl)- 1H-pyrazol-3-yl)-pyrimidin-4(3H)-one as the starting pyrimidone.

Stage 3: Synthesis of reference compound I-236

The title compound was prepared following general procedure B, except that (S)-3-methyl-2-(methylamino)butanoic acid was the amine reagent, 2-(3-(4-chloro-5-) was used fluoropyrimidin-2-yl)-1-(2-fluorobenzyl)-1H-pyrazol-5-yl)oxazole instead of intermediate 1, and the contents were heated at 100 °C for 41 h. The crude material was purified by silica gel chromatography (0-20% gradient (acetonitrile/methanol=7:1) in dichloromethane) to provide the desired compound, reference compound I-236 (8.9 mg, 49%). during two stages) as a whitish solid.

1H-NMR (400 MHz, CDCh) 88.25 (d, 1H), 7.70 (s, 1H), 7.46 (s, 1H), 7.24 (s, 1H), 7.21 (m 1H), 7.07-6.95 (m, 3H), 6.11 (d, 1H), 6.04 (d, 1H), 4.27 (d, 1H), 3.23 (d, 3H ), 2.52 (m, 1H), 1.11 (d, 3H), 0.94 (d, 3H).

Reference compound I-36

To a stirred solution of the reference compound I-12 dissolved in dichloromethane, an equivalent volume of trifluoacetic acid was added at 23 °C. The contents were stirred until all starting material consumption was observed by LC/MS. The reaction was diluted with dichloromethane and quenched with saturated sodium bicarbonate solution. The layers were separated, and the organic layer was washed with saturated sodium bicarbonate solution, water, and brine. The organic layer was further dried over Na2SO4, filtered and concentrated in vacuo to provide the desired compound, reference compound I-36 (19.5 mg, 75% yield) as a solid.

1H NMR (500 MHz, CDCla) 88.46 (s, 1 H), 8.15 (d, 1 H), 7.27 - 7.30 (m, 1 H), 7.13 - 7.23 ( m, 1 H), 7.00 - 7.09 (m, 1 H), 6.91 -7.00 (m, 1 H), 6.81 -6.90(m, 1 H), 6, 54-6.62 (m, 1 H), 5.95 (s, 2 H), 5.19 (d, 1 H), 4.26 - 4.40 (m, 1 H), 3.23 - 3.35 (m, 2 H), 3.03 (br. s, 1H), 2.92 (td, 2 H), 2.10-2.20 (m, 2 H), 1.59 - 1 .77 (m, 2 H).

Reference compound I-37

An equivalent volume of trifluoacetic acid was added to a stirred solution of the reference compound I-25 dissolved in dichloromethane at 23 °C. The contents were stirred until all starting material consumption was observed by LC/MS. The reaction was diluted with dichloromethane and quenched with saturated sodium bicarbonate solution. The layers were separated, and the organic layer was washed with saturated sodium bicarbonate solution, water, and brine. The organic layer was further dried over Na2SO4, filtered and concentrated in vacuo to provide the desired compound, reference compound I-37 (14 mg, 79% yield) as a solid.

1H NMR (500 MHz, CDCta) 88.45 (d, 1 H), 8.16 (d, 1 H), 7.30 (s, 1 H), 7.14 - 7.23 (m, 1 H) ), 7.00 - 7.07 (m, 1 H), 6.96 (td, 1 H), 6.78 - 6.89 (m, 1 H), 6.60 (d, 1 H), 5.96 (s, 2 H), 4.39 - 4.51 (m, 1 H), 4.22 - 4.36 (m, 1 H), 3.25 (ddd, 1 H), 2, 93-3.08 (m, 2H), 1.99-2.11 (m, 1 H), 1.83- 1.92 (m, 1 H), 1.59- 1.72 (m, 1 H), 1.37- 1.50 (m, 1 H).

Reference compound I-38

To a stirred solution of the reference compound I-26 dissolved in dichloromethane, an equivalent volume of trifluoacetic acid was added at 23 °C. The contents were stirred until all starting material consumption was observed by LC/MS. The reaction was diluted with dichloromethane and quenched with saturated sodium bicarbonate solution. The layers were separated, and the organic layer was washed with saturated sodium bicarbonate solution, water, and brine. The organic layer was further dried over Na2SO4, filtered and concentrated in vacuo to provide the desired compound, reference compound I-38 (11 mg, 55% yield) as a solid.

1H NMR (500 MHz, CDCta) 88.46 (d, 1 H), 8.16 - 8.22 (m, 1 H), 7.30 (m, 1 H), 7.17 - 7.25 ( m, 2 H), 7.00 - 7.09 (m, 1 H), 6.91 - 6.98 (m, 1 H), 6.85 (d, 1 H), 6.56 - 6, 68 (m, 1 H), 5.96 - 6.03 (m, 1 H), 5.95 (s, 2 H), 4.94 - 5.01 (m, 1 H), 4.39 ( t, 2H), 3.99 (dd, 2H).

REFERENCE COMPOSITION I-39

To a stirred solution of the reference compound I-27 dissolved in dichloromethane, an equivalent volume of trifluoacetic acid was added at 23 °C. The contents were stirred until all starting material consumption was observed by LC/MS. The reaction was diluted with dichloromethane and quenched with saturated sodium bicarbonate solution. The layers were separated, and the organic layer was washed with saturated sodium bicarbonate solution, water, and brine. The organic layer was further dried over Na2SO4, filtered and concentrated in vacuo to provide the desired compound, reference compound I-39 (12.2 mg, 69% yield) as a solid.

1H NMR (500 MHz, CDCla) 88.45 (d, 1H), 8.13 (d, 1 H), 7.35 (s, 1 H), 7.14-7.24 (m, 1 H) , 6 .99-7.08 (m, 1 H), 6.92 6.99 (m, 1 H), 6.79 - 6.89 (m, 1 H), 6.64 (d, 1 H ), 5.97 (s, 2 H), 5.55 (br. s., 1 H), 4.34 - 4.49 (m, 1 H), 3.29 (dd, 1 H), 2 .88-3.01 (m, 1 H), 2.79-2.87 (m, 1 H), 2.75(dd, 1 H), 1.90-2.02 (m, 1 H) , 1.77- 1.88 (m, 1 H), 1.58 - 1.76 (m, 2 H).

Reference compound I-40

An equivalent volume of trifluoacetic acid was added to a stirred solution of the reference compound I-21 dissolved in dichloromethane at 23 °C. The contents were stirred until all starting material consumption was observed by LC/MS. The reaction was diluted with dichloromethane and quenched with saturated sodium bicarbonate solution. The layers were separated, and the organic layer was washed with saturated sodium bicarbonate solution, water, and brine. The organic layer was further dried over Na2SO4, filtered and concentrated in vacuo to provide the desired compound, reference compound I-40 (22 mg, 90% yield) as a solid.

1H NMR (500 MHz, methanol-d4) 88.73 (d, 1 H), 8.06 (d, 1 H), 7.37 (s, 1 H), 7.19-7.30 (m, 1 H), 7.04-7.11 (m, 1 H), 7.00 (t, 1 H), 6.86 (d, 1 H), 6.77 (t, 1 H), 5, 93 (s, 2 H), 4.58 (t, 2 H), 4.13 (dd, 2 H), 3.98 - 4.09 (m, 1 H).

Reference compound I-133

An equivalent volume of trifluoacetic acid was added to a stirred solution of the reference compound I-101 dissolved in dichloromethane at 23 °C. The contents were stirred until all starting material consumption was observed by LC/MS. The mixture was concentrated in vacuo and the resulting gum was triturated with diethyl ether, filtered, and the solids were washed with diethyl ether. The solids were collected and dried under vacuum to provide the desired compound, reference compound I-133 (as TFA salt, 100 mg, 83% yield) as a white solid.

1H NMR (500 MHz, DMSO-de) 89.11 (s, 1 H), 8.87 (s, 1 H), 8.47 (d, 1 H), 7.58 (s, 1 H), 7.29 - 7.40 (m, 1 H), 7.17 -7.27 (m, 2 H), 7.11 (t, 1 H), 6.83 (t, 1 H), 5, 89 (s, 2 H), 5.44-5.49 (m, 1 H), 4.59 - 4.64 (m, 1 H), 3.79 (d, 1 H), 3.41 - 3.46 (m, 1 H), 3.11-3.18 (m, 1 H), 3.01 -3.12 (m, 1 H).

Reference compound I-30

The title compound was prepared following general procedure B, except that 2,8-diazaspiro[4.5]decane-3-one (2 equiv.) was the amine reagent, no triethylamine was used, and the contents were heated to 40 °C for 2 d as a solution in THF. The reaction was cooled, the solvent was removed in vacuo and the resulting solid was rinsed with 1N HCl solution to provide the desired compound, reference compound I-30 (57.8 mg, 83% yield) as a white solid.

1H NMR (500 MHz, METHANOL-d4) 8 ppm 8.86 (s, 1 H) 8.36 (d, 1 H) 7.71 (s, 1 H) 7.31 - 7.37 (m, 1 H) 7.13 (dd, 2 H) 6.99 - 7.04 (m, 2 H) 6.06 (s, 2 H) 4.32 (br. s., 2 H) 4.13 (br s., 2 H) 3.37 (br. s., 2 H) 2.42 (s, 2 H) 1.95 (t, 4 H).

Reference compound I-42

The title compound was prepared following general procedure B, except that 2-oxa-7-azaspiro[3.5]nonane oxalate (2 equiv.) was the amine reagent, and 8 equivalents of triethylamine were used, and the content was heated to 40 °C for 24 h as a solution in NMP. The reaction was cooled, diluted with ethyl acetate, and the mixture was rinsed with water. The contents were dried, filtered and concentrated in vacuo. The crude material was purified by silica gel chromatography with a gradient of 0-80% ethyl acetate/hexanes providing the desired compound, reference compound I-42 (42 mg, 52% yield) as a white solid.

1H NMR (500 MHz, METHANOL-d4) 8 ppm 8.75 (s, 1 H) 8.13 (d, 1 H) 7.41 (s, 1 H) 7.20 - 7.32 (m, 1 H) 6.97 - 7.14 (m, 2 H) 6.90 (s, 1 H) 6.81 (t, 1 H) 5.95 (s, 2 H) 4.52 (s, 4 H) ) 3.80 - 3.88 (m, 4 H) 1.90 - 2.05 (m, 4 H).

Reference compound I-43

The title compound was prepared following general procedure B, except that 8-oxa-2-azaspiro[4.5]decane (2 equiv.) was the amine reagent, no triethylamine was used, and the contents were heated to 40° C for 24 h as a solution in THF. The reaction was cooled and concentrated to give a solid, which was dissolved in ethyl acetate. The organic layer was washed with 1N HCl aqueous solution, dried, filtered and concentrated in vacuo. The crude material was purified by silica gel chromatography with a gradient of 0-100% ethyl acetate/hexanes providing the desired compound, reference compound I-42 (6.4 mg, 17% yield) as a white solid. .

1H NMR (500 MHz, METHANOL-d4) 5 ppm 8.77 (s, 1 H) 8.09 (d, 1 H) 7.44 (s, 1 H) 7.25 - 7.33 (m, 1 H) 7.11 (t, 1 H) 7.05 (s, 1 H) 6.93 (s, 1 H) 6.82 (s, 1 H) 5.98 (s, 2 H) 3.92 (br. s., 2 H) 3.71 - 3.84 (m, 6 H) 2.00 (t, 2 H) 1.64 - 1.75 (m, 4 H).

Reference compound I-32

The title compound was prepared following general procedure B, except that 2-oxa-6-azaspiro[3.3]heptane oxalate (2 equiv.) was the amine reagent, and 6 equivalents of triethylamine were used, and the content was heated to 40 °C for 2 h as a solution in<t>H<f>. The reaction was cooled and concentrated to give a solid, which was dissolved in ethyl acetate. The organic layer was washed with 1N HCl solution, dried, filtered and concentrated in vacuo to provide the desired compound, reference compound I-32 (19 mg, 33% yield) as a white solid.

1H NMR (500 MHz, METHANOL-d4) 5 ppm 8.83 (s, 1 H) 8.24 (br. s, 1 H) 7.57 (br. s., 1 H) 7.29 - 7, 33 (m, 1 H) 7.03 -7.16 (m, 2 H) 6.91 - 7.01 (m, 2 H) 5.99 - 6.05 (m, 2 H) 4.28 - 4.61 (m, 4 H) 3.99 (s, 2 H) 3.88 (s, 2 H).

Reference compound I-47

The title compound was prepared following general procedure B, except that 2-oxa-6-azaspiro[3.5]nonane oxalate (2 equiv.) was the amine reagent, and 4 equivalents of triethylamine were used, and the content was heated to 40 °C for 2 h as a solution in NMP. The reaction was cooled and concentrated to give a solid, which was dissolved in ethyl acetate. The organic layer was washed with water, dried, filtered and concentrated in vacuo. The crude material was purified by silica gel chromatography with a gradient of 0-100% ethyl acetate/hexanes providing the desired compound, reference compound I-47 (42.3 mg, 66% yield) as a white solid. .

1H NMR (400 MHz, METHANOL-d4) 5 ppm 8.67 - 8.76 (m, 1 H) 8.12 (d, 1 H) 7.40 (s, 1 H) 7.19 - 7.28 (m, 1 H) 7.02 -7.10 (m, 1 H) 6.99 (t, 1 H) 6.88 (d, 1 H) 6.79 (t, 1 H) 5.89 - 5.95 (m, 2 H) 4.38 - 4.49 (m, 4 H) 4.05 - 4.10 (m, 2 H) 3.71 - 3.79 (m, 2 H) 1, 92 - 1.98 (m, 2 H) 1.58 - 1.68 (m, 2 H).

Reference compound I-44

The title compound was prepared following general procedure B, except that fed-butyl 2,8-diazaspiro[4.5]decane-2-carboxylate (2 equiv.) was the amine reagent, no triethylamine was used, and the content was heated to 40 °C for 2 d as a solution in THF. The reaction was cooled and diluted with ethyl acetate. The organic layer was washed with water, dried, filtered and concentrated in vacuo. The crude material was purified by silica gel chromatography with a gradient of 0-100% ethyl acetate/hexanes providing the desired compound, reference compound I-44 (51.6 mg, 67% yield) as a white solid. .

1H NMR (500 MHz, METHANOL-d4) 5 ppm 8.77 (s, 1 H)8.15(d, 1 H) 7.43 (s, 1 H) 7.23 - 7.33 (m, 1 H) 7.11 (t, 1 H) 7.05 (t, 1 H) 6.92 (s, 1 H) 6.83 (t, 1 H) 5.97 (s, 2 H) 3.79 - 4.09 (m, 4 H) 3.43 - 3.52 (m, 2 H) 3.30 (s, 2 H) 1.89 (t, 2 H) 1.72 (br. s., 4H) 1.49 (s, 9H).

Reference compound I-45

The title compound was prepared following general procedure B, except that fed-butyl 2,7-diazaspiro[4.4]nonane-2-carboxylate (2 equiv.) was the amine reagent, 1 equivalent of triethylamine was used, and the content was heated to 40 °C for 2 d as a solution in THF. The reaction was cooled and diluted with ethyl acetate. The organic layer was washed with water, dried, filtered and concentrated in vacuo. The crude material was purified by silica gel chromatography with a gradient of 0-100% ethyl acetate/hexanes providing the desired compound, reference compound I-45 (48.3 mg, 64% yield) as a white solid. .

1H NMR (500 MHz, METHANOL-d4) 5 ppm 8.75 (s, 1 H) 8.04 - 8.13 (m, 1 H) 7.40 (s, 1 H) 7.24 - 7.32 (m, 1 H) 7.10 (t, 1 H) 7.03 (t, 1 H) 6.89 (s, 1 H) 6.82 (t, 1 H) 5.96 (s, 2 H) ) 3.85 - 3.98 (m, 2 H) 3.69 - 3.82 (m, 2 H) 3.49 (br. s., 2 H) 3.30 -3.43 (m, 4 H) 1.48 (d, 9 H).

Reference compound I-61

The title compound was prepared following general procedure B, except that 3,3-difluoroazetidine (as the HCl salt, 1 equiv.) was the amine reagent, 2 equivalents of Hunig's base were used in place of triethylamine, and the content was heated at 40 °C for 3 h as a solution in NMP. The reaction was cooled and diluted with ethyl acetate. The organic layer was washed with water, dried, filtered and concentrated in vacuo. The crude material was purified by silica gel chromatography with a gradient of 0-30% ethyl acetate/hexanes providing the desired compound, reference compound I-61 (37 mg, 71% yield) as a white solid.1

1H NMR (400 MHz, CDCla) 5 ppm 8.44 (d, 1 H) 8.22 (d, 1 H) 7.30 (s, 1 H) 7.14 - 7.21 (m, 1 H) 6.91 - 7.04 (m, 2 H) 6.81 (t, 1 H) 6.54 - 6.59 (m, 1 H) 5.95 (s, 2 H) 4.60 - 4. 71 (m, 4 H).

I-62 REFERENCE COMPOSITION

The title compound was prepared following general procedure B, except that 4,4-difluoropiperidine (as the HCl salt, 1 equiv.) was the amine reagent, 2 equivalents of Hunig's base were used in place of triethylamine, and the content was heated at 40 °C for 3 h as a solution in NMP. The reaction was cooled and diluted with ethyl acetate. The organic layer was washed with water, dried, filtered and concentrated in vacuo. The crude material was purified by silica gel chromatography with a gradient of 0-30% ethyl acetate/hexanes providing the desired compound, reference compound I-62 (40.4 mg, 71% yield) as a white solid. . 1H NMR (400 MHz, CDCla) 8 ppm 8.43 (d, 1 H) 8.22 (d, 1 H) 7.24 - 7.25 (m, 1 H) 7.12 - 7.21 (m , 1 H) 6.91 - 7.04 (m, 2 H) 6.82 (t, 1 H) 6.56 (d, 1 H) 5.94 (s, 2 H) 3.94 - 4, 02 (m, 4 H) 2.04 - 2.17 (m, 4 H).

Reference compound I-63

The title compound was prepared following general procedure B, except that 3,3-difluoro-pyrrolidine (as HCl salt, 1 equiv.) was the amine reagent, 2 equivalents of Hunig's base were used instead of triethylamine , and the content was heated at 40 °C for 3 h as a solution in NMP. The reaction was cooled and diluted with ethyl acetate. The organic layer was washed with water, dried, filtered and concentrated in vacuo. The crude material was purified by silica gel chromatography with a gradient of 0-30% ethyl acetate/hexanes providing the desired compound, reference compound I-63 (41.5 mg, 71% yield) as a white solid. .

1H NMR (400 MHz, CDCla) 8 ppm 8.43 (d, 1 H) 8.19 (d, 1 H) 7.29 (s, 1 H) 7.11 - 7.22 (m, 1 H) 6.90 - 7.04 (m, 2 H) 6.78 - 6.87 (m, 1 H) 6.56 (d, 1 H) 5.94 (s, 2 H) 3.98 - 4. 18 (m, 4 H) 2.40 - 2.54 (m, 2 H).

Reference compound I-166

The title compound was prepared following general procedure B, except that A/-benzylglycine ethyl ester (1 equiv.) was the amine reagent, 2 equivalents of triethylamine were used, and the contents were heated to 80 °C for 24 h as a solution in THF/water (10:1). The reaction was cooled and diluted with ethyl acetate. The organic layer was washed with saturated ammonium chloride solution, dried, filtered and concentrated in vacuo. The crude material was purified by silica gel chromatography to provide the desired compound, reference compound I-166 (33 mg, 47%) as a white solid.

1H NMR (400 MHz, CDCh) 8 ppm 8.42 (d, 1 H) 8.23 (d, 1 H) 7.28 - 7.39 (m, 5 H) 7.23 (d, 1 H) 7.12 - 7.21 (m, 1 H) 6.91 - 7.05 (m, 2 H) 6.83 (t, 1 H) 6.53 (d, 1 H) 5.94 (s, 2 H) 5.00 (s, 2 H) 4.20 - 4.24 (m, 2 H) 4.14 - 4.20 (m, 2 H) 1.21 (t, 3 H).

Reference compound I-167

The title compound was prepared following general procedure B, except that ethyl W-methylaminoacetate (as the HCl salt, 1 equiv.) was the amine reagent, 2 equivalents of triethylamine were used, and the contents were heated to 90 °C for 24 h as a solution in THF. The reaction was cooled and diluted with ethyl acetate and water. The layers were separated and the organic layer was dried, filtered and concentrated in vacuo. The crude material was purified by silica gel chromatography with a gradient of 0-100% ethyl acetate/hexanes providing the desired compound, reference compound I-167 (77 mg, 79% yield) as a white solid.

1H NMR (500 MHz, CDCh) 8 ppm 8.46 (d, 1 H) 8.22 (d, 1 H) 7.28 (d, 1 H) 7.15 - 7.25 (m, 1 H) 6.95 - 7.06 (m, 2 H) 6.81 - 6.89 (m, 1 H) 6.58 (d, 1 H) 5.95 - 6.00 (m, 2 H) 4. 35 (s, 2 H) 4.23 (q, 2 H) 3.43 (d, 3 H) 1.25 (t, 3 H).

Reference compound I-176

A mixture of reference compound I-167 (70 mg, 1 equiv.) and sodium hydroxide [1.0 N aqueous solution] (770 pl, 5 equiv.) in THF (385 pl) and MeOH (385 pl) was stirred at rt for 24 h. The mixture was quenched with 1N HCl (5 equiv.). The white precipitate formed was collected by filtration, clarified with a minimum amount of ether and dried to give the reference compound I-176 (52 mg, 79% yield) as a white solid.

1H NMR (500 MHz, DMSO-d6) 8 ppm 9.11 (d, 1 H) 8.34 (d, 1 H) 7.54 (s, 1 H) 7.30-7.37 (m, 1 H) 7.19-7.25 (m, 2 H) 7.11 (t, 1 H) 6.86 (t, 1 H) 5.90 (s, 2 H) 4.41 -4.45 ( m, 2 H) 3.32 (d, 3 H).

Reference compound I-168

A mixture of reference compound I-167 (30 mg, 1 equiv) and sodium hydroxide [1.0 N aqueous solution] (57 pl, 1 equiv) in THF (141 pl) and MeOH (141 pl) was stirred. at rt for 24 h. The mixture was treated with 1N HCl (5 equiv.). The mixture was diluted in dichloromethane (100 ml) and washed with water (50 ml). The organic layer was dried, filtered and evaporated to give an oil. The oil was purified by column chromatography (0 to 10% methanol in dichloromethane) giving the reference compound I-168 (10 mg, 36% yield) as a white solid.1

1H NMR (400 MHz, METHANOL-d4) 8 ppm 8.74 (d, 1 H) 8.20 (d, 1 H) 7.33 - 7.43 (m, 5 H) 7.22 - 7.32 (m, 3 H) 6.99 -7.13 (m, 3 H) 6.84 (d, 2 H) 5.95 (s, 2 H) 5.05 (s, 2 H).

I-218 REFERENCE COMPOSITION

A mixture of reference compound I-176 (48 mg, 1 equiv.), O-methylhydroxylamine hydrochloride (14 mg, 1.5 equiv.), EDC (32 mg, 1.5 equiv.) and DMAP (21 mg, 1.5 equiv) in DMF (563 jl) was stirred at rt for 2 h. The mixture was diluted in ethyl acetate (100 ml) and washed with water (50 ml x 3). The organic layer was dried, filtered and evaporated to give an oil. The oil was purified by column chromatography giving the reference compound I-218 (26 mg, 51% yield) as a white solid.

1H NMR (500 MHz, METHANOL-d4) 8 ppm 8.63 - 8.68 (m, 1 H) 8.08 (d, 1 H) 7.30 (s, 1 H) 7.13 - 7.20 (m, 1 H) 6.96 -7.02 (m, 1 H) 6.92 (t, 1 H) 6.77 (s, 1 H) 6.71 (t, 1 H) 5.85 ( s, 2 H) 4.17 (s, 2 H) 3.55 (s, 3 H) 3.30 - 3.34 (m, 3 H).

Reference compound I-223

The title compound was prepared following general procedure B, except that W-methyl-1-(3-methyl-1,2,4-oxadiazol-5-yl)methanamine (as HCl salt, 1 equiv.) was amine reagent, 4 equivalents of triethylamine were used, and the content was heated at 85 °C for 24 h as a solution in dioxane. The reaction was cooled and diluted with ethyl acetate. The organic layer was washed with 1N HCl solution, dried, filtered and concentrated in vacuo. The crude material was purified by silica gel chromatography with a gradient of 0-100% ethyl acetate/hexanes providing the desired compound, reference compound I-223 (16.3 mg, 21% yield) as a white solid. .

1H NMR (500 MHz, CDCh) 8 ppm 8.46 (d, 1 H) 8.25 (d, 1 H) 7.15 - 7.22 (m, 2 H) 6.96 (t, 1 H) 6.84 (t, 1 H) 6.61 (d, 1 H) 5.95 (s, 2 H) 5.03 (s, 2 H) 3.52 (d, 3 H) 2.38 (s , 3H).

Reference compound I-14

2-(1-(2-fluorobenzyl)-5-(isoxazol-3-yl)-1H-pyrazol-3-yl)-5-nitropyrimidin-4-ol (1 equiv.) (this intermediate was found in a previous patent: WO2012/3405 A1) (25 mg, 1 equiv.) was treated with POCh (457 μl, 75 equiv.) and stirred at reflux for 1.5 h. The content was concentrated in vacuo and the residue was azeotroped with toluene (2 times). The residue was redissolved in THF (0.7 ml) and treated with morpholine (171 μl, 30 equiv.). The contents were heated to 40 °C, and the reaction was stirred for 1.5 h. The residue was transferred to a 1:1 mixture of ethyl acetate and water. The layers were separated and the aqueous layer was extracted with ethyl acetate (3 times). The organic parts were combined and washed with brine. The mixture was dried over MgSO4, filtered and concentrated in vacuo to give the desired reference compound I-14 (30 mg, 97%) as a pale yellow solid.

1H-NMR (400 MHz, CDCla) 88.47 (d, 1 H), 8.36 (d, 1 H), 8.09 - 8.16 (m, 1 H), 7.69 (dd, 1 H), 7.41 (d, 1 H), 7.20 (t, 1 H), 6.66 - 6.70 (m, 1 H), 6.45 (d, 1 H), 6.06 (s, 2 H), 3.79 - 3.86 (m, 4 H), 3.74 (m, 4 H).

Reference compound I-15

A solution of reference compound I-14 (30 mg, 1 equiv) in methanol was treated with palladium on carbon (7 mg, 10 wt % palladium, 0.1 equiv) and placed under a hydrogen atmosphere. The contents were stirred for 2 h at 23 °C. The content was filtered over celite and eluted with methanol. The contents were concentrated in vacuo and the crude material was purified by silica gel chromatography using a gradient of 0-70% (acetonitrile:methanol=7:1)/dichloromethane providing the desired reference compound I-15 (11.5 mg, 39%) as a white solid. 1H-NMR (400 MHz, CDCh) 88.43 (d, 1 H), 8.36 (d, 1 H), 7.35 (d, 1 H), 6.80 (t, 1 H), 6 .59-6.53 (m, 1 H), 6.49-6.40 (m, 2 H), 6.11 (dd, 1 H), 5.93-5.82 (m, 2 H) , 3.87-3.76 (m, 4 H), 3.72 (d, 4 H).

Reference compound I-70

A solution of reference compound I-37 in toluene was treated with ethyl isocyanate (3 equiv.) and heated at 90 °C for 20 min. The resulting precipitates were filtered and rinsed with toluene. The solids were collected and dried under vacuum to provide the desired compound, reference compound I-70 (7 mg, 36% yield) as a solid.

1H NMR (500 MHz, DMSO-de) 89.09 (d, 1 H), 8.27 (d, 1 H), 7.77 (s, 1 H), 7.30 - 7.36 (m, 1 H), 7.14 - 7.27 (m, 2 H), 7.07 - 7.12 (m, 1 H), 6.83 (t, 1 H), 5.94 (d, 1 H ), 5.91 (s, 2 H), 5.76 (t, 1 H), 4.17 (d, 1 H), 3.94 (d, 1 H), 3.69 (dt, 1 H ), 3.52 (t, 1 H), 3.22 (dd, 1 H), 3.02 (quin, 2 H), 1.85 (d, 1 H), 1.71 -1.81 ( m, 1 H), 1.43 - 1.62 (m, 2 H), 0.97 (t, 3 H).

Reference compound I-71

A solution of reference compound I-40 in toluene was treated with ethyl isocyanate (3 equiv.) and heated at 90 °C for 20 min. The resulting precipitates were filtered and rinsed with toluene. The solids were collected and dried under vacuum to provide the desired compound, reference compound I-71 (3 mg, 16% yield) as a solid.

1H NMR (500 MHz, DMSO-de) 89.10 (d, 1 H), 8.25 (d, 1 H), 7.49 - 7.55 (m, 1 h) 7.29 - 7. 37 (m, 1 H), 7.19 - 7.27 (m, 2 H), 7.10 (t, 1 H), 6.81 (t, 1 H), 6.52 - 6.61 ( m, 1 H), 6.02 (t, 1 H), 5.90 (s, 2 H), 4.51 - 4.59 (m, 1 H), 4.47 (m, 2 H), 4.06 (d, 2 H), 2.94 - 3.07 (m, 2 H), 0.99 (t, 3 H).

REFERENCE COMPOSITION I-136

A solution of reference compound I-133 in dichloromethane was treated with ethyl isocyanate (1.1 equiv.) and triethylamine (2 equiv.), and stirred at 23 °C for 1 h. The solvent was removed in vacuo and the residue was resuspended in diethyl ether. The resulting precipitates were filtered and rinsed with diethyl ether. The solids were collected and dried under vacuum to provide the desired compound, reference compound I-136 (2.9 mg, 28% yield) as a solid.

1H NMR (500 MHz, DMSO-de) 8 9.09 (s, 1 H), 8.26 - 8.39 (m, 1 H), 7.52 (s, 1 H) 7.32 (m, 1 H), 7.17 - 7.27 (m, 2 H), 7.10 (t, 1 H), 6.78-6.88 (m, 1 H), 6.63 (br.s. , 1 H), 5.90 (m, 2 H), 4.90 - 5.19 (m, 1 H) 4.43 (d, 1 H), 4.30 (br.s., 1 H) , 3.91 (d, 1 H), 3.48 (d, 1 H), 3.22 (d, 1 H), 2.99 - 3.05 (m, 3 H), 1.01 (t , 3H).

Reference compound I-134

A solution of reference compound I-133 in dichloromethane was treated with propionyl chloride (1.1 equiv.) and triethylamine (2 equiv.), and stirred at 23 °C for 1 h. The solvent was removed in vacuo and the residue was resuspended in diethyl ether. The resulting precipitates were filtered and rinsed with diethyl ether. The solids were collected and dried under vacuum to provide the desired compound, reference compound I-134 (3.5 mg, 35% yield) as a solid.

1H NMR (500 MHz, DMSO-de) 8 9.10 (s, 1 H), 8.40 (d, 1 H), 7.49 - 7.64 (m, 1 H), 7.28 - 7.39 (m, 1 H), 7.15 - 7.26 (m, 2 H), 7.11 (t, 1 H), 6.84 (br. s., 1 H), 5.89 (s, 2 H), 4.79 (d, 1 H), 4.24 - 4.45 (m, 2 H), 3.91 (d, 1 H), 3.67 (br. s., 1 H) , 3.58 (d, 1 H), 2 .87-3.00 (m, 1 H), 2.31 -2.40 (m, 2 H), 0.93- 1.04 (m, 3 H).

Reference compound I-135

A solution of reference compound I-133 in dichloromethane was treated with methyl chloroformate (1.1 equiv.) and triethylamine (2 equiv.), and stirred at 23 °C for 1 h. The solvent was removed in vacuo and the residue was resuspended in diethyl ether. The resulting precipitates were filtered and rinsed with diethyl ether. The crude material was purified by reverse phase HPLC providing the desired compound, reference compound I-135 (3.5 mg, yield

37%) as a solid.

1H NMR (500 MHz, CD3OD) 88.82 (s, 1 H), 8.41 (d, 1 H), 7.58 (s, 1 H), 7.26 - 7.35 (m, 1 H ), 7.12 (t, 1 H), 7.07 (t, 1 H), 6.88 - 6.97 (m, 2 H), 6.01 (s, 2 H), 5.51 ( br. s., 1 H), 4.68 (d, 1 H), 4.58 (br. s., 1 H), 4.09 (d, 1 H), 3.79 - 3.95 ( m, 1 H), 3.76 (s, 3 H), 3.52 - 3.62 (m, 1 H), 3.35 - 3.45 (m, 1 H).

Reference compound I-49 and reference compound I-50

A solution of 1-methylcyclopropanecarboxylic acid (141 mg, 10 equiv.) in dichloromethane (1 ml) was treated with oxalyl chloride (0.11 ml, 9 equiv.), and the contents were stirred until no further bubbling was observed. . The resulting solution was then added little by little over 5 minutes to a cooled (0 °C) solution of 5-fluoro-2-(1-(2-fluorobenzyl)-5-(isoxazol-3-yl)-1H- pyrazol-3-yl)-pyrimidin-4-amine (1 equiv, 50 mg) this intermediate is described in a previous patent application publication, WO2012/3405 A1) in dichloromethane (0.35 ml) and pyridine (0 .35ml). The mixture was heated to 60 °C and stirred for 24 h. The reaction mixture was diluted with ethyl acetate and washed with saturated aqueous ammonium chloride solution. The layers were separated and the aqueous layer was extracted with ethyl acetate (3 times) and dichloromethane (1 time). The organic parts were combined and washed with brine. The mixture was dried over MgSO4, filtered and concentrated in vacuo. The crude material was purified by silica gel chromatography using a gradient of 0-60% ethyl acetate/hexanes providing the desired reference compound I-49 (18.5 mg, 30%) as a white solid, along with I -50 (16.2 mg, 22%) as a clear oil.

Reference compound I-49 H1 NMR (400 MHz, CDCh) 88.57 (d, 1 H), 8.43 (d, 1 H), 8.01 (s, 1 H), 7.38 (s, 1 H), 7.22-7.13 (m, 1 H), 7.00 (t, 1 H), 6.98-6.90 (m, 1 H), 6.78 (t, 1 H) ), 6.57 (d, 1 H), 5.99 (s, 2 H), 1.48 (s, 3 H), 1.38-1.32

(m, 2 H), 0.79-0.73 (m, 2 H).

Reference compound I-50 H1 NMR (400 MHz, CDCla) 88.64 (d, 1 H), 8.47-8.44 (m, 1 H), 7.28 (s, 1 H), 7. 22-7,15

(m, 1 H), 7.02 (d, 1 H), 6.97 (t, 1 H), 6.93-6.87 (m, 1 H), 6.55-6.53 (m , 1 H), 5.95 (s, 2 H), 1.53-1.48 (m, 4 H), 1.22 (s, 6 H), 0.85-0.79 (m, 4 H).

Reference compound I-51 and reference compound I-52

A solution of 5-fluoro-2-(1-(2-fluorobenzyl)-5-(isoxazol-3-yl)-1H-pyrazol-3-yl)-pyrimidin-4-amine (1 equiv, this intermediate in a previous patent application publication WO2012/3405 A1) (see above) (50 mg, 1 equiv.) in THF (0.7 ml) was cooled to 0 ° C and treated with LiHMDS (0.16 ml) , 1.1 equiv., 1 M solution) and stirred for 20 minutes. The reaction was then treated with methyl chloroformate (44 pL, 4 equiv.). The reaction mixture was stirred at 0°C for 20 minutes, then heated to 23°C for 1 hour. The reaction was diluted with ethyl acetate and quenched with saturated aqueous ammonium chloride solution. The layers were separated and the aqueous layer was extracted with ethyl acetate (twice) and dichloromethane (three times). The organic parts were combined and washed with brine. The mixture was dried over MgSO4, filtered and concentrated in vacuo. The crude material was purified by silica gel chromatography using a gradient of 0-100% ethyl acetate/hexanes providing the desired reference compound I-51 (5.3 mg, 9%) as an off-white solid, along with the reference compound I-52 (13.1 mg, 20%) as a white solid.

Reference compound I-51 1H-NMR (500 MHz, CDCh) 88.56 (s, 1 H), 8.45 (s, 1 H), 7.41 (s, 1 H), 7.38 (s , 1 H), 7.19 (m, 1 H), 7.02 (t, 1 H), 6.95 (m, 1 H), 6.81 (m, 1 H), 6.61 (s , 1 H), 6.00 (s, 2 H), 3.87 (s, 3 H).

Reference compound I-521H-NMR (500 MHz, CDCh) 88.77 (s., 1 H), 8.47 (s, 1 H), 7.40 (s, 1 H), 7.21 (m , 1 H), 7.07-6.93 (m, 2 H), 6.84 (m, 1 H), 6.59 (s, 1 H), 6.02 (s, 2 H), 3 .83 (s, 6 H).

Reference compound I-144

In a small vial, reference compound I-58 (0.022 g, 0.047 mmol) was diluted with DCM (volume: 2.0 ml) then filled with CDI (28 mg, 0.173 mmol). The reaction mixture was then heated to 45 °C until complete consumption of the starting acid was verified by LC/MS. At this time, cyclopropanesulfonamide (22.86 mg, 0.189 mmol) and DBU (7.11 μL, 0.047 mmol) were added and the reaction was heated for an additional 30 minutes. At that time, the reaction was quenched with 1N HCl, then diluted with DCM. The layers were separated and the aqueous part was extracted two more times with DCM. The organic parts were combined, dried (Na2SO4), filtered, and concentrated. The crude material was purified using SiO2 chromatography with a gradient of 0-10% MeOH/DCM providing the desired sulfonamide, reference compound I-144 (16 mg, 54% yield).

1H-NMR (400 MHz, CDCla) 810.62 (bs, 1H), 8.43 (d, 1H), 8.23 (d, 1H), 7.45 (s, 1H), 7.19 (dd , 1H), 7.03-6.95 (m, 2H), 6.85 (t, 1H), 6.69 (s, 1H), 5.96 (dd, 2H), 4.20-4, 12 (m, 1H), 2.87-2.79 (m, 1H), 2.30-2.24 (m, 1H), 2.02-1.92 (m, 1H), 1.86- 1.70 (m, 4H), 1.30-0.86 (m, 6H).

Reference compound I-157

The title compound was prepared using the procedure described for reference compound I-144, except for the use of reference compound I-88 as the starting carboxylic acid. Purification by silica gel chromatography provided the desired compound, reference compound I-157 (10 mg, 55% yield) as a solid.

1H-NMR (400 MHz, CD3OD) 88.76 (d, 1H), 8.18 (d, 1H), 7.55 (s, 1H), 7.25 (dd, 1H), 7.07 (t , 1H), 7.00 (t, 1H), 6.92 (d, 1H), 6.81 (t, 1H), 5.96 (dd, 2H), 4.66-4.62 (m, 1H), 2.88-2.83 (m, 1H), 1.93-1.83 (m, 2H), 1.31-1.27 (m, 2H), 1.16-1.10 ( m, 1H), 1.04 (d, 3H), 0.97 (d, 3H), 0.92-0.78 (m, 2H).

Reference compound I-187

The title compound was prepared using the procedure described for reference compound I-144, except for the use of reference compound I-89 as the starting carboxylic acid. Purification by silica gel chromatography provided the desired compound, reference compound I-187 (33 mg, 80% yield) as a solid.

1H-NMR (400 MHz, DMSO-de) 811.92 (s, 1H), 9.09 (d, 1H), 8.33 (d, 1H), 7.55 (s, 1H), 7.29 (dd, 1H), 7.18 (t, 1H), 7.12 (d, 1H), 7.07 (t, 1H), 6.84 (t, 1H), 5.87 (s, 2H) , 4.50 (d, 1H), 3.19 (d, 3H), 2.95-2.87 (m, 1H), 2.42-2.35 (m, 1H), 1.05 (d , 3H), 1.02-0.94 (m, 2H), 0.89 (d, 3H), 0.87-0.83 (m, 2H).

Reference compound I-272

The title compound was prepared following general procedure B, except that (S)-3-methyl-2-(methylamino)butanoic acid was the amine reagent, 3-(3-(4-chloropyrimidin-2-) was used. il)-1-(2-fluorobenzyl)-1H-pyrazol-5-yl)isoxazole (synthesis described in the procedure for reference compound I-24) instead of intermediate I and the contents were heated to 110 °C for 72 hours. The crude material was purified by silica gel chromatography with a gradient of 0-10% methanol/dichloromethane providing the desired compound, reference compound I-272 (4 mg, 8% yield) as a solid.

1H-NMR (400 MHz, DMSO-de) 812.98 (bs, 1H), 9.07 (s, 1H), 8.24 (bs, 2H), 7.48 (bs, 1H), 7.30 (dd, 1H), 7.19 (t, 1H), 7.07 (t, 1H), 6.84 (bs, 1H), 6.60 (bs, 1H), 5.86 (s, 2H) , 5.24 (bs, 1H), 2.94 (bs, 3H), 2.30 (bs, 1H), 1.02 (d, 3H), 0.77 (d, 3H).

Reference compound I-74

Intermediate 1 was dissolved in THF and cooled to 0 °C. In a separate vial, 1H-pyrazole (1 equiv.) was diluted with THF, then sodium hydride (60% in dispersion oil, 1 equiv.) was introduced to generate the sodium salt. The content was allowed to stir for 15 min. At that time, the sodium salt was added portionwise to the solution of intermediate 1. Once the starting material was consumed as observed by LC/MS, the reaction was quenched with 1N HCl and the mixture was extracted with dichloromethane (three times). The organic portions were combined, filtered and concentrated. The crude material was then purified using a gradient of 0-10% methanol/dichloromethane providing the desired compound, reference compound I-74 (41 mg, 72% yield).

1H-NMR (400 MHz, DMSO-da) 89.10 (t, 1H), 9.03 (dd, 1H), 8.80-8.79 (m, 1H), 8.02-8.00 ( m, 1H), 7.81 (d, 1H), 7.34 7.28 (m, 1H), 7.24 (t, 1H), 7.23-7.18 (m, 1H), 7, 09 (t, 1H), 6.88 (t, 1H), 6.72-6.70 (m, 1H), 5.93 (s, 2H).

Reference compound I-273

This compound was synthesized according to general procedure B using 2-((2,2,2-trifluoroethyl)amino)acetic acid hydrochloride. After completing the consumption of starting material, the solution was diluted with aqueous 1N sodium hydroxide to pH ~ 10. Diethyl ether was added and the layers were separated. The aqueous layer was acidified with aqueous hydrochloric acid to pH ~ 2. Ethyl acetate was added, and the layers were separated again. The aqueous layer was extracted with ethyl acetate and the combined organic materials were dried over magnesium sulfate, filtered, and the solvent was removed in vacuo. Purification by silica gel chromatography (0-15% methanol in dichloromethane) provided the reference compound I-273 (6 mg, 23%) as a white solid.

1H-NMR (400 MHz, CD3OD) 88.73 (m, 1H), 8.27 (d, 1H), 7.40 (s, 1H), 7.27-7.22 (m, 1H), 7 .09-6.99 (m, 2H), 6.87-6.86 (m, 1H), 6.80 (t, 1H), 5.94 (s, 2H), 4.61-4.55 (m, 2H), 4.45 (s, 2H).

Reference compound I-274

The title compound was prepared following general procedure B, except that 3-((methylamino)methyl)benzoic acid (as the HCl salt) was the amine reagent, and the contents were heated at 90 °C for 2 h as a solution in dioxane. Ethyl acetate was used as the extraction solvent during processing. The crude compound of reference compound I-274 (20 mg, 68% yield) was isolated as a white solid for which no further purification was needed.

1H-NMR (400 MHz, CD3OD) 88.76 (m, 1H), 8.29 (d, 1H), 8.08 (s, 1H), 7.97 (d, 1H), 7.65-7 .63 (m, 1H), 7.54 (s, 1H), 7.48 (t, 1H), 7.30-7.24 (m, 1H), 7.10-7.01 (m, 2H ), 6.94-6.88 (m, 2H), 5.99 (s, 2H), 5.16 (s, 2H), 3.48 (d, 3H).

Reference compound I-275

The title compound was prepared following general procedure B, except that 4-((methylamino)methyl)benzoic acid (as the HCl salt) was the amine reagent. Ethyl acetate was used as the extraction solvent during processing. The crude compound of reference compound I-275 (17 mg, 63% yield) was isolated as a white solid for which no further purification was needed.

1H-NMR (400 MHz, DMSO-de) 8 12.88 (br s, 1H), 9.05 (m, 1H), 8.25 (d, 1H), 7.87 (d, 2H), 7 .48 (s, 1H), 7.44 (d, 2H), 7.32-7.27 (m, 1H), 7.21-7.19 (m, 2H), 7.07 (t, 1H ), 6.83 (t, 1H), 5.86 (s, 2H), 4.95 (s, 2H), 3.24 (d, 3H).

Reference compound I-276

This compound was synthesized according to general procedure B using 1 H-tetrazol-5-amine and dioxane as a solvent. The crude residue was suspended in dichloromethane and filtered. The filtrate was purified by silica gel chromatography (0-10% methanol in dichloromethane) to provide the reference compound I-276 (0.4 mg, 2% yield) as a white film.

1H-NMR (400 MHz, CD3OD) 88.75 (m, 1H), 8.42 (d, 1H), 7.48 (s, 1H), 7.29-7.24 (m, 1H), 7 .11-7.06 (m, 1H), 7.05-7.01 (m, 1H), 6.89-6.83 (m, 2H), 5.97 (s, 2H).

Reference compound I-277

This compound was synthesized according to general procedure B using 3-amino-3-methylbutanoic acid and the contents were heated at 80 °C for 68 h. Purification by silica gel chromatography (0-10% methanol in dichloromethane) provided the reference compound I-277 (1.3 mg, 5% yield) as a white film.

1H-NMR (400 MHz, CD3OD) 88.74 (m, 1H), 8.02 (d, 1H), 7.34 (d, 1H), 7.27-7.22 (m, 1H), 7 .09-7.00 (m, 2H), 6.89-6.83 (m, 2H), 5.93 (s, 2H), 3.03 (s, 2H), 1.66 (s, 6H) ).

Reference compound I-278

This compound was synthesized according to general procedure B using 5-(aminomethyl)pyridin-2(1 H)-one and the content was stirred at 90 °C for 40 h. The crude reaction mixture was diluted with 1N aqueous hydrochloric acid and ethyl acetate. The layers were separated and the aqueous layer was concentrated in vacuo. Purification by reverse phase HPLC (gradient 20-50% acetonitrile in water with 0.1% TFA, 20 min) provided the reference compound I-278 (13 mg, 35% yield) as a tan solid. .1

1H-NMR (400 MHz, CD3OD) 88.82 (s, 1H), 8.28 (d, 1H), 7.73-7.64 (m, 3H), 7.32-7.26 (m, 1H), 7.11-7.03 (m, 2H), 6.99 6.95 (m, 2H), 6.54 (d, 1H), 6.01 (s, 2H), 4.73 ( s, 2H).

REFERENCE COMPOSITION I-279

This compound was synthesized according to general procedure B using the trifluoroacetic acid salt of 2-((methylamino)methyl)benzoic acid with dioxane as a solvent and the contents were heated at 90 °C for 2 days. Purification of the crude reaction mixture by reverse phase HPLC (gradient of 5-75% acetonitrile in water with 0.1% TFA, 20 min) gave the reference compound I-279 (15 mg, 37% yield). like a white solid.

1H-NMR (400 MHz, DMSO-de) 813.17 (br s, 1H), 9.10 (m, 1H), 8.25 (d, 1H), 7.94 (d, 1H), 7, 56-7.52 (m, 1H), 7.44 (s, 1H), 7.41-7.29 (m, 3H), 7.24-7.19 (m, 2H), 7.12- 7.08 (m, 1H), 6.85-6.81 (m, 1H), 5.89 (s, 2H), 5.24 (s, 2H), 3.30 (s, 3H).

Reference compound I-280

This compound was synthesized according to general procedure B using 4-(aminomethyl)benzoic acid with ethyl acetate as extraction solvent. Purification of the crude reaction mixture by reverse phase HPLC (gradient 5-75% acetonitrile in water with 0.1% TFA, 20 min) gave the reference compound I-280 (3.4 mg, yield 9 %) as a white solid.

1H-NMR (400 MHz, CD3OD) 88.81 (s, 1H), 8.26 (d, 1H), 8.02 (d, 2H), 7.58 (d, 2H), 7.53 (s , 1H), 7.33-7.27 (m, 1H), 7.13-7.04 (m, 2H), 6.95-6.91 (m, 2H), 6.01 (s, 2H) ), 5.01 (s, 2H).

Reference compound I-281

This compound was synthesized according to general procedure B using 6-methylpiperidine-2-carboxylic acid. Purification of the crude reaction mixture by silica gel chromatography (0-10% methanol in dichloromethane) provided the reference compound I-281 (3.4 mg, 9% yield) as a white solid.

1H-NMR (400 MHz, CD3OD) 88.75 (d, 1H), 8.15 (d, 1H), 7.40 (s, 1H), 7.29-7.23 (m, 1H), 7 .11-7.06 (m, 1H), 7.02 (td, 1H), 6.88 (d, 1H), 6.82-6.78 (m, 1H), 5.95 (s, 2H) ), 5.46 (br s, 1H), 2.46-2.43 (m, 1H), 1.91-1.72 (m, 4H), 1.63-1.60 (m, 2H) , 1.35 (d, 3H).

Reference compounds I-282 and I-283

These were synthesized according to general procedure B using a mixture of (1R,4S)-4-methylpiperidine-2-carboxylic acid and (1S,4S)-4-methylpiperidine-2-carboxylic acid. Purification of the crude reaction mixture by silica gel chromatography (0-10% methanol in dichloromethane) provided the reference compound I-282 (15 mg, 39% yield) as a white solid. Repurification of the mixed fractions by reverse phase HPLC (5-75% acetonitrile in water with 0.1% TFA) provided the reference compound I-283 (4 mg, 10% yield).

Reference compound I-282 1H-NMR (400 MHz,CD3OD) 88.74 (m, 1H), 8.22 (d, 1H), 7.41 (s, 1H), 7.26-7.21 ( m, 1H), 7.08-7.04 (m, 1H), 7.01-6.98 (m, 1H), 6.84 (m, 1H), 6.81-6.78 (m, 1H), 5.93 (s, 2H), 4.44 (dd, 1H), 4.04-3.98 (m, 1H), 3.65-3.60 (m, 1H), 2.19 (dt, 1H), 1.93-1.70 (m, 3H), 1.46-1.38 (m, 1H), 1.04 (d, 3H).

Reference compound I-283 1H-NMR (400 MHz, CD3OD) 88.77 (d, 1H), 8.28 (d, 1H), 7.52 (s, 1H), 7.29-7.23 ( m, 1H), 7.10-7.00 (m, 2H), 6.92 (d, 1H), 6.88-6.85 (m, 1H), 5.97 (s, 2H), 5 .68 (br s, 1H), 4.74 (br s, 1H), 3.41 (br s, 1H), 2.44-2.39 (m, 1H), 1.87-1.82 ( m, 1H), 1.74-1.65 (m, 1H), 1.58-1.50 (m, 1H), 1.38-1.28 (dq, 1H), 1.00 (d, 3H).

Reference compound I-237

This compound was synthesized according to general procedure B using (R)-W,2-dimethyl-1-(1H-tetrazol-5-yl)propan-1-amine (2 equivalents). Purification of the crude reaction mixture by reverse phase HPLC (gradient of 5-75% acetonitrile in water with 0.1% TFA) provided the reference compound I-237 (4 mg, 23% yield) as an oil. transparent.

1H-NMR (400 MHz, CD3OD) 88.80 (m, 1H), 8.31 (d, 1H), 7.53 (s, 1H), 7.28-7.25 (m, 1H), 7 .09-7.01 (m, 3H), 6.96 (m, 1H), 6.05 (d, 1H), 5.98 (d, 1H), 5.76 (br s, 1H), 3 .35 (d, 3H), 2.86-2.80 (m, 1H), 1.07 (d, 3H), 0.90 (d, 3H).

Reference compound I-284

This compound was synthesized according to general procedure B using (R)-2-methyl-1-(1H-tetrazol-5-yl)propan-1-amine. Purification of the crude reaction mixture by silica gel chromatography (0-10% methanol in dichloromethane) provided the reference compound I-284 (16 mg, 37% yield) as a white solid.

1H-NMR (400 MHz, CD3OD) 88.75 (m, 1H), 8.12 (d, 1H), 7.27-7.21 (m, 2H), 7.06-7.03 (m, 1H), 7.01-6.98 (m, 1H), 6.88 (d, 1H), 6.82-6.78 (m, 1H), 5.96 (d, 1H), 5.91 (d, 1H), 5.50 (d, 1H), 2.61-2.52 (m, 1H), 1.14 (d, 3H), 0.93 (d, 3H).

I-285 REFERENCE COMPOSITION

To a solution of reference compound I-147 (previously described, 1 equivalent) and pyridine (50 equivalents) in dichloromethane at 0 °C was added cyclopropanecarbonyl chloride (1.2 equivalents) over 30 seconds. The solution was immediately warmed to room temperature and stirred for a further 2.5 hours. After diluting with saturated aqueous ammonium chloride and dichloromethane, the layers were separated and the aqueous layer was extracted with dichloromethane. The organic substances were dried over magnesium sulfate, filtered, and the solvent was removed in vacuo. Purification by silica gel chromatography (0-5% methanol in dichloromethane) provided the reference compound I-285 (11 mg, 34% yield) as a white solid.

1H-NMR (400 MHz, CD3OD) 88.73 (m, 1H), 8.18 (d, 1H), 7.43 (s, 1H), 7.27-7.22 (m, 1H), 7 .09-7.04 (m, 1H), 7.00 (t, 1H), 6.90 (m, 1H), 6.77 (t, 1H), 5.95 (s, 2H), 3, 98-3.95 (m, 4H), 3.46-3.44 (m, 4H), 1.65-1.59 (m, 1H), 0.92-0.84 (m, 4H).

Reference compound I-229

The title compound was prepared following general procedure B, except that 2-(piperidin-3-yl)acetic acid was the amine reagent, 6 equivalents of Hunig's base were used instead of triethylamine, and the contents were heated up to 120 °C for 18 h as a solution in THF/water (10:1). The solvent was removed under a stream of nitrogen. and the crude material was purified by reverse phase HPLC to provide the desired compound, reference compound I-229 (8.1 mg, 32% yield) as a solid.

1H NMR (500 MHz, CD3OD) 8 ppm 8.84 (m, 1 H), 8.28 (m, 1 H), 7.73 (m, 1 H), 7.32 (m, 1 H), 7.12 (m, 2 H), 6.98 (m, 2 H), 6.04 (s, 2 H), 4.96 (m, 1 H), 4.67 (m, 1 H), 3.54 (m, 1 H), 3.27 (m, 1 H), 2.42 (m, 2 H), 2.25 (m, 1 H), 2.00 (m, 2 H), 1.79 (m, 1 H), 1.54 (m, 1 H).

Reference compound I-230 and reference compound I-231

The title compound was prepared following general procedure B, except that a mixture of 2-(piperidin-4-yl)acetic acid and methyl 2-(piperidin-4-yl)acetate was the amine reagent, 6 equivalents of Hunig base instead of triethylamine, and the content was heated to 120 °C for 18 h as a solution in THF/water (10:1). The solvent was removed under a stream of nitrogen, and the resulting crude material was purified by reverse phase HPLC to provide the desired compounds, the reference compound I-230 (6.5 mg, 25% yield) as a solid, and the Reference compound I-231 (16.2 mg, 61% yield) as a solid.

1H NMR for Reference Compound I-230 (500 MHz, M ETAN O L^) 8 ppm 8.83 (m, 1 H), 8.26 (m, 1 H), 7.63 (m, 1 H) , 7.30 (m, 1 H), 7.10 (m, 2 H), 6.97 (m, 2 H), 6.03 (s, 2 H), 4.98 (m, 2 H) , 3.40 (m, 1 H), 2.35 (m, 2 H), 2.25 (m, 1 H), 2.04 (m, 2 H), 1.50 (m, 2 H) .

1H NMR for Reference Compound I-231 (500 MHz, M ETAN O L^) 8 ppm 8.84 (m, 1 H), 8.30 (m, 1 H), 7.66 (m, 1 H) , 7.28-7.37 (m, 1 H), 7.05-7.17 (m, 2 H), 7.00 (d, 2 H), 6.04 (s, 2 H), 4 .93-5.02 (m, 2 H), 3.70 (s, 3 H), 3.35-3.45 (m, 2H), 2.358 (d, 2 H), 2.22-2, 34 (m, 1 H), 1.99-2.08 (m, 2 H), 1.50 (br.s., 2 H).

Reference compound I-232

The title compound was prepared following general procedure B, except that 2-amino-4-methoxybutanoic acid was the amine reagent, 6 equivalents of Hunig's base were used instead of triethylamine, and the contents were heated to 120° C for 18 h as a solution in THF/water (10:1). The solvent was removed under a stream of nitrogen and the crude material was purified by reverse phase HPLC to provide the desired compound, reference compound I-232 (10 mg, 40% yield) as a solid.

1H NMR (500 MHz, CD3OD) 8 (ppm) 8.80 (m, 1 H), 8.32 (m, 1 H), 7.55 (s, 1 H), 7.29 (m, 1 H) ), 7.08 (m, 2 H), 6.96 (m, 2 H), 6.01 (s, 2 H), 5.11 (m, 1 H), 3.61 (m, 2 H ), 3.35 (s, 3 H), 2.43 (m, 1 H), 2.21 (m, 1 H).

Reference compound I-234

The title compound was prepared following general procedure B, except that 3-(piperidin-4-yl)propanoic acid was the amine reagent. The solvent was removed under a stream of nitrogen and the crude material was purified by reverse phase HPLC to provide the desired compound, reference compound I-234 (13 mg, 49% yield) as a solid.

1H NMR (500 MHz, CD3OD) 8 (ppm) 8.81 - 8.86 (m, 1 H), 8.25 - 8.31 (m, 1 H), 7.61 - 7.67 (m, 1 H), 7.29 - 7.36 (m, 1 H), 7.05-7.16 (m, 2 H), 6.95-7.02 (m, 2 H), 6.04 ( s, 2 H), 4.93 - 5.02 (m, 2 H), 3.37 (s, 2 H), 2.36-2.45 (m, 2 H), 1.96 - 2, 06 (m, 2 H), 1.76 - 1.88 (m, 1 H), 1.61 - 1.70 (m, 2 H), 1.35 - 1.48 (m, 2 H).

Reference compound I-286

The title compound was prepared following the general procedure B, except that 4-(aminomethyl)phenol was the reagent of am ina (1.1 equiv.), 4 equiva lents of triethylamine were used, and the contents were heated to 90 °C for 12 h as or a solution in dioxane/water (10:1). The crude material was purified by silica gel chromatography with a gradient of 1-5% methanol/dichloromethane over 40 min providing the desired compound, reference compound I-286 (17.7 mg, 48% yield) as a solid. white.

1H NMR (500 MHz, CDCh) 8 (ppm): 8.45 (d, 1H), 8.16 (d, 1H), 7.34 (s, 1H), 7.16- 7.24 (m, 3H), 6.99 - 7.04 (m, 1H), 6.94 - 6.99 (m, 1H), 6.85 - 6.90 (m, 1H), 6.79 - 6.83 (m, 2H), 6.58 (d, 1H), 5.97 (s, 2H), 5.67 (br. s, 1H), 5.28 - 5.30 (m, 1H), 4.72 (d , 2H).

Reference compound I-287

The title compound was prepared following general procedure B, except that (4-(methylsulfonyl)phenyl)methanamine was the amine reagent (1 equiv.), 4 equivalents of triethylamine were used, and the contents were heated to 90° C for 12 h as a solution in dioxane/water (10:1). The crude material was purified by silica gel chromatography with a gradient of 1-5% methanol/dichloromethane over 40 min providing the desired compound, reference compound I-287 (23.3 mg, 56% yield) as a solid. white.

1H NMR (500 MHz, CDCla) 8 (ppm): 8.47 (d, 1H), 8.22 (m, 1H), 7.90 - 7.96 (m, 2H), 7.64 - 7, 68 (m, 2H), 7.20 - 7.25 (m, 2H), 6.98 - 7.08 (m, 2H), 6.88 - 6.93 (m, 1H), 6.57 ( d, 1H), 5.98 (s, 2H), 5.52 - 5.63 (br. d, 1H), 4.93 - 4.97 (m, 2H), 3.06 (s, 3H) .

Reference compound I-288

The title compound was prepared following general procedure B, except that 2-(aminomethyl)phenol was the amine reagent (1.1 equiv.), 4 equivalents of triethylamine were used, and the contents were heated to 90°C. for 12 h as a solution in dioxane/water (10:1). The crude material was purified by silica gel chromatography with a gradient of 1-5% methanol/dichloromethane over 40 min providing the desired compound, reference compound I-288 (4.5 mg, 12% yield) as a solid. white.

1H NMR (500 MHz, CDCh) 8 (ppm): 9.52 (s, 1H), 8.49 (d, 1H), 8.15 (d, 1H), 7.42 (s, 1H), 7 .21 - 7.27 (m, 3H), 7.02 -7.11 (m, 3H), 6.89 - 6.95 (m, 2H), 6.64 (d, 1H), 6.03 (s, 2H), 5 .80 -5 .85 (m, 1H), 4.75 (d, 2H).

Reference compound I-289

The title compound was prepared following general procedure B, except that 2-(4-methylpiperidin-4-yl)acetic acid was the amine reagent, 4 equivalents of triethylamine were used, and the contents were heated to 90°C. for 12 h as a solution in dioxane/water (3:1). A mixture of dichloromethane/isopropanol (5:1) was used as extraction solvent. The crude material was purified by silica gel chromatography with a gradient of 1-5% methanol/dichloromethane over 40 min providing the desired compound, reference compound I-289 (37.4 mg, 70% yield) as a solid. sparkling white.

1H NMR (500 MHz, CDCh) 8 (ppm): 8.45 (s, 1H), 8.12 (d, 1H), 7.30 (s, 1H), 7.16 - 7.22 (m, 1H), 7.00 - 7.06 (m, 1H), 6.94 - 6.98 (m, 1H), 6.82 - 6.88 (m, 1H), 6.59 (d, 1H), 5 .97 (s, 2H), 3.83 - 3.96 (m, 2H), 3.59 (s, 2H), 2.42 - 2.49 (m, 2H), 1.76- 1.86 (m, 4H), 1.15 (s, 3H).

Reference compound I-290

The title compound was prepared following general procedure B, except that 4-cyclohexylpiperidine-4-carboxylic acid (as the TFA salt, 1.2 equiv.) was the amine reagent, 4 equivalents of triethylamine were used, and the content was heated at 90 °C for 12 h as a solution in dioxane/water (3:1). A mixture of dichloromethane/isopropanol (5:1) was used as extraction solvent. The crude material was purified by silica gel chromatography with a 1-5 methanol/dichloromethane gradient over 40 min providing the desired compound, reference compound I-290 (44.6 mg, 76% yield) as a solid. white.

1H NMR (500 MHz, CDCh) 8 (ppm): 8.47 (s, 1H), 8.17 (d, 1H), 7.31 (s, 1H), 7.16 - 7.24 (m, 1H), 7.01 - 7.08 (m, 1H), 6.95 - 7.00 (m, 1H), 6.83 - 6.88 (m, 1H), 6.60 (d, 1H), 5 .99 (s, 2H), 4.58 - 4.65 (m, 2H), 3.07 - 3.18 (m, 2H), 2.24 -2.32 (m, 2H), 1.77 - 1.86 (m, 4H), 1 .45 - 1.70 (m, 3H), 1 .13 -1 .26 (m, 3H), 1 .03 -1 .13 (m, 3H).

Reference compound I-291

The title compound was prepared following general procedure B, except that methyl 2-phenylpiperidine-2-carboxylate was the amine reagent, 4 equivalents of sodium bicarbonate were used instead of triethylamine, and the contents were heated to 110 °C for 48 h. Ethyl acetate was used as the extraction solvent. First pass purification was achieved by silica gel chromatography using 1-5% methanol in dichloromethane gradient over 40 minutes providing the product at 80% purity. Further purification was achieved using reverse phase HPLC using a gradient of 5-95% acetonitrile in water over 30 minutes providing the analytically pure desired compound, reference compound I-291 (2 mg, 3% yield) as a white solid.

1H NMR (500 MHz, CDCI3) 5 (ppm): 8.41 (s, 1H), 8.35 (br. s, 1H), 7.24-7.27 (m, 2H), 7.09 - 7 .18 (m, 3H), 7 .02 -7 .09 (m, 1H), 6.90 - 7.00 (m, 2H), 6.79 - 6.87 (m, 1H), 6, 67 - 6.74 (m, 1H), 6.44 - 6.51 (m, 1H), 5.86 (d, 1H), 5.74 (d, 1H), 3.72 - 3.85 ( m, 1H), 3.36 - 3.51 (m, 1H), 2.47 - 2.56 (m, 1H), 1.70 - 1.99 (m, 5H).

Reference compound I-292

The title compound was prepared following general procedure B, except that 4-amino-2-phenylbutanoic acid (as the HCl salt) was the amine reagent, 4 equivalents of triethylamine were used, and the contents were heated to 95 °C for 12 h. Ethyl acetate was used as the extraction solvent. The crude material was purified by silica gel chromatography with a gradient of 1-5% methanol in dichloromethane over 40 minutes providing the desired compound, reference compound I-292 (37.9 mg, 50% yield) as a solid. white.

1H NMR (500 MHz, CDCh) 5 (ppm): 8.47 (d, 1H), 8.13 (d, 1H), 7.29 - 7.36 (m, 6H), 7.15 - 7, 23 (m, 1H), 6.99 - 7.04 (m, 1H), 6 .92-6.97 (m, 1H), 6 .86 -6 .91 (m, 1H), 6.60 ( d, 1H), 6.01 (d, 1H), 5.94 (d, 1H), 5.21 -5 .28 (m, 1H), 3 .85 3.94 (m, 1H), 3, 62 - 3.80 (m, 2H), 2.51 -2 .61 (m, 1H), 2.11 -2 .19 (m , 1H).

Reference compound I-293

The title compound was prepared following general procedure B, except that 4-methoxypiperidine-4-carboxylic acid (as the TFA salt) was the amine reagent (2 equiv.), 4 equivalents of triethylamine were used, and the content was heated to 105 °C as a solution in dioxane/water (3:1) for 12 h. Ethyl acetate was used as the extraction solvent. The crude material was purified by silica gel chromatography using a gradient of 1 to 5% methanol in dichloromethane over 40 minutes providing the desired compound, reference compound I-293 (52.7 mg, 56% yield) as a solid. white.

1H NMR (500 MHz, CDCla) 5 (ppm): 8.47 (s, 1H), 8.22 (d, 1H), 7.31 (s, 1H), 7.16 - 7.25 (m, 1H), 7.01 - 7.09 (m, 1H), 6.95 - 7.01 (m, 1H), 6.84 - 6.89 (m, 1H), 6.60 (d, 1H) , 5.98 (s, 2H), 4.33 - 4.41 (m, 2H), 3.53 - 3.62 (m, 2H), 3.41 (s, 3H), 2.05-2 .20 (m, 4H).

Reference compound I-294

The title compound was prepared following general procedure B, except that 2-(piperidin-4-yl)propanoic acid was the amine reagent (2 equiv.), 4 equivalents of triethylamine were used, and the contents were heated to 90 °C for 12 h as a solution in dioxane/water (3:1). A mixture of dichloromethane/isopropanol (5:1) was used as extraction solvent. The crude material was purified by silica gel chromatography with a gradient of 1 to 5% methanol in dichloromethane over 40 minutes providing the desired compound, reference compound I-294 (42.8 mg, 68% yield) as an off-white solid. .

1H NMR (500 MHz, CDCh) 5 (ppm): 8.47 (s, 1H), 8.20 (d, 1H), 7.31 (s, 1H), 7.17 - 7.23 (m, 1H), 7.00 - 7.07 (m, 1H), 6.94 - 7.02 (m, 1H), 6.81 - 6.89 (m, 1H), 6.60 (d, 1H) , 5.98 (s, 2H), 4.70 - 4.84 (m, 2H), 3.01 - 3.06 (t, 2H), 2.39 - 2.44 (m, 1H), 1 .93 - 2.01 (m, 1H), 1.82 - 1.93 (m, 2H), 1.37 - 1.54 (m, 2H), 1.24 (d, 3H).

Reference compound I-295

The title compound was prepared following general procedure B, except that 4-phenylpiperidine-2-carboxylic acid (as the TFA salt) was the amine reagent (2 equiv.), 4 equivalents of triethylamine were used, and the content was heated at 110 °C for 64 h. Ethyl acetate was used as the extraction solvent. The crude material was purified by silica gel chromatography with a gradient of 1 to 5% methanol in dichloromethane over 40 minutes providing the desired compound, reference compound I-295 (12.0 mg, 18% yield) as a mixture. racemic with cis configuration (as a whitish solid).

1H NMR (500 MHz, CD3OD) 5 (ppm): 8.76 (s, 1H), 8.21 (d, 1H), 7.45 (s, 1H), 7.25 - 7.36 (m, 5H), 7.19 -7.25 (m, 1H), 7.08 - 7.14 (m, 1H), 7.02 - 7.07 (m, 1H), 6.91 (d, 1H) , 6.80 - 6.86 (m, 1H), 5.97 (s, 2H), 5.62 - 5.77 (m, 1H), 2.77 -2.89 (m, 1H), 2 .55 -2 .62 (m, 1H), 2 .03 -2 .12 (m, 1H), 1 .96 -2 .02 (m, 1H), 1 .83 - 1.96 (m, 1H) , 1 .25 - 1.35 (m, 1H), 0.84-0.98 (m, 1H).

Reference compound I-296

The title compound was prepared following general procedure B, except that 4-(4-methoxyphenyl)piperidine-4-carboxylic acid (as the TFA salt) was the amine reagent (2 equiv.), 4 equivalents were used of triethylamine, and the content was heated at 110 °C for 17 h as a solution in dioxane/water (3:1). Ethyl acetate was used as the extraction solvent. The crude material was purified by silica gel chromatography using a gradient of 1 to 5% methanol in dichloromethane over 40 minutes providing the desired compound, reference compound I-296 (41.1 mg, 66% yield) as a solid. white.

1H NMR (400 MHz, CDCh) 5 (ppm): 8.43 (s, 1H), 8.12 (d, 1H), 7.34 (d, 2H), 7.28 (s, 1H), 7 .12 - 7.16 (m, 1H), 6.94 -7.02 (m, 1H), 6.88 - 6.93 (m, 1H), 6.86 (d, 2H), 6.75 - 6.80 (m, 1H), 6.56 (d, 1H), 5.94 (s, 2H), 4.44 - 4.52 (m, 2H), 3.78 (s, 3H), 3.36 - 3.41 (m, 2H), 2.63 - 2.72 (m, 2H), 1.96 - 2.08 (m, 2H).

I-298 REFERENCE COMPOSITION

The title compound was prepared following general procedure B, except that 4-aminopiperidine-4-carboxylic acid (as the HCl salt) was the amine reagent, and the contents were heated at 100 °C for 18 h as a solution in THF/DMF/triethylamine (1:1:1). After complete consumption of the starting material, the reaction was cooled to 0° and an excess of 2 M trimethylsilyldiazomethane solution was added and stirred at 23 °C for 3 d until the conversion to the amine ester was complete. The contents were diluted with 1N NaOH solution and extracted with dichloromethane. The organic layer was dried, filtered and concentrated in vacuo. The crude material was purified by silica gel chromatography with a gradient from 20 to 100% ethyl acetate in hexanes providing the desired compound, reference compound I-298 (22 mg, 63% yield) as a white solid.

1H NMR (500 MHz, DMSO-da) 8 ppm 9.08 (d, 1 H), 8.28 (d, 1 H), 7.54 (s, 1 H), 7.29 - 7.39 ( m, 1 H), 7.18 - 7.27 (m, 2 H), 7.10 (t, 1 H), 6.83 (t, 1 H), 5.90 (s, 2 H), 4.02 - 4.09 (m, 2 H), 3.66 - 3.74 (m, 2 H), 3.63 - 3.65 (m, 3 H), 1.99 - 2.04 ( m, 2 H), 1.91 -1.98 (m2 H), 1.62 (d, 2 H).

Reference compound I-299

The title compound was prepared following general procedure B, except that 4-aminopiperidine-4-carboxylic acid (as the HCl salt) was the amine reagent (5 equiv.), 8 equivalents of triethylamine were used, and the Content was heated at 90 °C for 18 h as a solution in THF/water (5:1). After complete consumption of the starting material, the reaction was cooled and filtered. The resulting crude material was collected and purified by reverse phase HPLC providing the desired compound, reference compound I-299 (2 mg, 7% yield) as a solid resultant.

1H NMR (500 MHz, METHANOL-d4) 8 ppm 8.80 (d, 1 H), 8.31 (d, 1 H), 7.54 (s, 1 H), 7.23 - 7.35 ( m, 1 H), 7.08 - 7.15 (m, 1 H), 7.05 (t, 1 H), 6.92 (d, 1 H), 6.81 - 6.90 (m, 1 H), 5.99 (s, 2 H), 4.45 (dt, 2 H), 3.96 - 4.13 (m, 2 H), 2.44 (dt, 2 H), 2, 02 (ddd, 2 H).

Reference compound I-300

The title compound was prepared following general procedure B, except that 4-hydroxypiperidine-4-carboxylic acid (as the HCl salt) was the amine reagent (5 equiv.), 8 equivalents of triethylamine were used, and the Content was heated at 90 °C as a solution in THF/water (5:1) for 18 h. After complete consumption of the starting material, the reaction was cooled and filtered. The filtrate was collected and concentrated in vacuo. The crude material was purified by reverse phase HPLC providing the desired compound, reference compound I-300 (22 mg, 81% yield) as a solid.

1H NMR (500 MHz, METHANOL-d4) 8 ppm 8.83 (d, 1 H), 8.22 - 8.35 (m, 1 H), 7.58 - 7.70 (m, 1 H), 7.25 - 7.37 (m, 1 H), 7.04-7.18 (m, 2 H), 6.90-7.02 (m, 2 H), 6.03 (s, 2 H) ), 4.76 (d, 2 H), 3.69-3.82 (m, 2 H), 2.16-2.33 (m, 2 H), 1.94 (d, 2 H).

Reference compound I-301

The title compound was prepared following general procedure B, except that (S)-4,4-difluoropyrrolidine-2-carboxylic acid was the amine reagent (5 equiv.), 8 equivalents of triethylamine were used, and the content was heated at 90 °C for 18 h as a solution in THF/water (5:1). After complete consumption of the starting material, the reaction was concentrated in vacuo. The crude material was purified by reverse phase HPLC providing the desired compound, reference compound I-301 (20 mg, 67% yield) as a solid.

1H NMR (500 MHz, CD3OD) 8 ppm 8.80 (d, 1 H), 8.34 (d, 1 H), 7.35 - 7.42 (m, 1 H), 7.24 - 7, 34 (m, 1 H), 7.10 (dd, 1 H), 7.01 - 7.07 (m, 1 H), 6.91 (td, 2 H), 5.98 (s, 2 H) ), 5.44 - 5.69 (m, 2 H) 4.76 - 4.87 (m, 3 H).

Reference compound I-302

The title compound was prepared following general procedure B, except that (S)-2-amino-3-ethoxypropanoic acid was the amine reagent (4 equiv.), 6 equivalents of triethylamine were used, and the contents were heated at 100 °C for 18 h as a solution in dioxane/water (3:1). After neutralization, the crude material was suspended in ethyl acetate and diluted with hexanes until precipitation occurred. The precipitate was filtered and collected to provide the desired compound, reference compound I-302 (9 mg, 24% yield) as a solid.

1H NMR (500 MHz, DMSO-d6) 8 ppm 9.08 (s, 1 H), 8.20 (d, 1 H), 7.47 (s, 1 H), 7.27 - 7.40 ( m, 2 H), 7.17 - 7.26 (m, 2 H), 7.10 (t, 1 H), 6.84 (t, 1 H), 5.81 - 5.98 (m, 2 H), 4.59 (br. s., 1 H), 3.83 - 3.90 (m, 1 H), 3.75 - 3.83 (m, 1 H), 3.45 -3 .54 (m, 1 H), 3.37 - 3.44 (m, 1 H), 0.92 - 1.08 (m, 3 H).

Reference compound I-303

The title compound was prepared by following the general procedure B, except that the acid 2-amin or -3-methoxipro Panic was the amine reagent (4 equiv.), 6 equiva lents of triethylamine were used, and the contents were heated to 100°. C for 18 h as a solution in dioxane/water (3:1). After neutralization, the crude material was suspended in ethyl acetate and diluted with hexanes until precipitation occurred. The precipitate was filtered and collected to provide the desired compound, reference compound I-303 (8 mg, 22% yield) as a solid.

1H NMR (500 MHz, DMSO-da) 8 ppm 9.09 (d, 1 H) 8.22 (d, 1 H), 7.47 (s, 1 H), 7.28 - 7.39 (m , 1 H), 7.17 - 7.27 (m, 2 H), 7.10 (t, 1 H), 6.85 (t, 1 H), 6.63 (br. s., 1 H ), 5.81 - 5.94 (m, 2 H), 4.54 - 4.88 (m, 1 H), 3.72 - 3.87 (m, 2 H), 3.57 (s, 2H), 3.25 (s, 3H).

Reference compound I-304

The title compound was prepared following step 3 of the procedure described for reference compound I-235, except that 1-((methylamino)methyl)cyclopropanecarboxylic acid (as the TFA salt) was the amine reagent, and the Contents were heated at 100 °C for 6 h. The crude material was purified by silica gel chromatography (1-4% methanol in dichloromethane gradient) providing the desired compound, reference compound I-304 (67 mg, 72% yield) as a white solid.

1H-NMR (400 MHz, CDCh) 88.77 (d, 1H), 8.10 (d, 1H), 7.39 (s, 1H), 7.16 (ap. q, 1H), 7.03 (ap. q, 1H), 6.92 (d, 1H), 6.68 (ap. t, 1H), 5.98 (s, 2H), 4.15 (s, 2H), 3.37 ( d, 3H), 1.28 (m, 2H), 1.07 (m, 2H).

Reference compound I-305

The title compound was prepared following step 3 of the procedure described for reference compound I-235, except that (2R,3S)-3-methylpiperidine-2-carboxylic acid (as the acetic acid salt) was the reagent. of amine, and the content was heated at 100 °C for 21 h. The crude material was purified by silica gel chromatography (2-4% methanol in dichloromethane gradient) providing the desired compound, reference compound I-305 (24 mg, 46% yield) as a white solid.

1H-NMR (400 MHz, CDCla) 88.77 (d, 1H), 8.21 (d, 1H), 7.47 (s, 1H), 7.15 (ap. q, 1H), 7.02 (ap. q, 1H), 6.89 (d, 1H), 6.66 (ap. t, 1H), 5.98 (s, 2H), 5.04 (d, 1H), 4.37 ( br. d, 1H), 3.70 (ap. t, 1H), 2.10 (m, 1H), 1.90 (br. d, 1H), 1.80-1.69 (m, 2H) , 1.52 (ap. q, 1H), 1.21 (d, 3H).

Reference compound I-306

The title compound was prepared following general procedure B, except that 4-isopropylpiperidine-4-carboxylic acid was the amine reagent, and the content was heated at 90 °C for 3 h as a solution in THF/water (10 :1). The contents were cooled to 23 °C, and the organic solvents were removed in vacuo. The solids were treated with 1 N HCl solution and the resulting precipitate was filtered and dried under vacuum to provide the desired compound, reference compound I-306 (42 mg, 86% yield) as a white solid.

1H-NMR (500 MHz, CDCh) 88.49 (d, 1 H), 8.35 (d, 1H), 7.64 (s, 1 H), 7.25-7.20 (m, 1 H ), 7.05-7.01 (m, 3 H), 6.67 (d, 1 H), 5.98 (s, 2 H), 4.80 (d, 2 H), 3.79- 3.72 (m, 1 H), 3.23 (t, 1 H), 2.35 (d, 2 H), 1.92-1.80 (m, 1 H), 1.62 (td, 1 H), 1.41 (t, 1 H), 0.97 (d, 6 H).

Reference compound I-307

The title compound was prepared following general procedure B, except that 3-(methylamino)bicyclo[1.1.1]pentane-1-carboxylic acid was the amine reagent, and the contents were heated at 90 °C for 18 h as a solution in THF/water (10:1). The crude material was purified by silica gel chromatography with a gradient of 0-10% methanol/dichloromethane providing the desired compound, reference compound I-307 (74 mg, 53% yield) as an off-white solid.

1H-NMR (500 MHz, CDCh) 88.53 (d, 1 H), 8.45 (d, 1 H), 7.49 (s, 1 H), 7.26-7.21 (m, 2 H), 7.09-7.01 (m, 2 H), 6.67 (d, 1 H), 5.93 (s, 2 H), 3.36 (d, 3 H), 2.68 (s, 6 H).

Reference compound I-308

The title compound was prepared following general procedure B, except that 2-azabicyclo[4.1.0]heptane-1-carboxylic acid (as the HCl salt) was the amine reagent, and the contents were heated to 90° C for 3 h as a solution in THF/water (10:1). The crude material was purified by silica gel chromatography with a gradient of 0-10% methanol/dichloromethane providing the desired compound, reference compound I-308 (32 mg, 17% yield) as an off-white solid.

1H-NMR (500 MHz, CDCh) 88.46 (d, 1 H), 8.21 (d, 1 H), 7.27 (s, 1 H), 7.23-7.18 (m, 1 H), 7.04 (t, 1 H), 6.98 (t, 1 H), 6.87 (t, 1 H), 6.59 (d, 1 H), 5.98 (s, 2 H), 4.62 (br. s., 1 H), 3.01 (br. s., 1 H), 2.20-2.11 (m, 1 H), 2.08-1.98 (m, 2 H), 1.83-1.72 (m, 2 H), 1.57-1.49 (m, 1 H), 1.04 (br. s., 1 H).

Reference compound I-309

The title compound was prepared following general procedure B, except that (1R,3S)-3-(Bocamino)cyclopentane-1-carboxylic acid (as the TFA salt) was the amine reagent, and the contents were heated at 90 °C for 3 h as a solution in THF/water (10:1). The crude material was purified by silica gel chromatography with a gradient of 0-10% methanol/dichloromethane providing an intermediate product. This intermediate product was immediately dissolved in THF, and cooled to 0 °C. The contents were treated with sodium hydride (60% in mineral oil, 2 equiv.) followed by methyl iodide (10 equiv.). The reaction was allowed to warm to 23 °C for 3 days. The content was poured onto water and extracted with ethyl acetate (3 times). The organic parts were combined and washed with brine. The mixture was dried over MgSO4, filtered and concentrated in vacuo. The crude material was purified by silica gel chromatography with a gradient of 0-15% methanol/dichloromethane providing the desired compound, reference compound I-309 (0.9 mg, 1% yield) as an off-white solid.

1H-NMR (500 MHz, CDCh) 88.45 (s, 1 H), 8.13 (d, 1 H), 7.32 (s, 1 H), 7.23-7.15 (m, 1 H), 7.03 (t, 1H), 6.97 (t, 1 H), 6.86 (t, 1 H), 6.60 (s, 1 H), 5.97 (s, 2 H) ), 4.75 (d, 1 H), 3.74 (s, 3 H), 3.08-2.94 (m, 1 H), 2.42-2.30 (m, 1 H), 2.17-1.84 (m, 5 H).

Reference compound I-310

The title compound was prepared following general procedure B, except that (2S, 3S)-2-methyl-piperidine-3-carboxylic acid was the amine reagent, and the contents were heated to 90 °C as a solution in THF/water (10:1) for 3 d. The crude material was purified by silica gel chromatography using a gradient of 0-50% (acetonitrile:methanol = 9:1 with 0.1% TFA)/dichloromethane providing the desired compound, reference compound I-310 ( 4.9 mg, 2% yield) as a white solid.

1H-NMR (500 MHz, CDCla) 88.46 (d, 1 H), 8.22 (d, 1 H), 7.31 (s, 1 H), 7.24-7.18 (m, 1 H), 7.13-7.01 (m, 1 H), 6.98 (t, 1 H), 6.87 (t, 1 H), 6.59 (d, 1 H), 5.97 (s, 2 H), 5.38 (br. s., 1 H), 4.42 (d, 1 H), 3.22 (t, 1 H), 2.99-2.87 (m, 1 H), 2.05 1.96 (m, 2 H), 1.93-1.84 (m, 2 H), 1.32 (d, 3 H).

Reference compound I-311

The title compound was prepared following general procedure B, except that (2R,3R)-2-methylpiperidine-3-carboxylic acid was the amine reagent, and the content was heated at 90 °C for 18 h as a solution. in THF/water (10:1). The crude material was purified by silica gel chromatography with a gradient of 0-10% methanol/dichloromethane providing the desired compound, reference compound I-311 (17.2 mg, 12% yield) as an off-white solid.

1H-NMR (500 MHz, DMSO-de) 812.51 (br. s., 1 H), 9.09 (d, 1 H), 8.31 (d, 1 H), 7.50 (s, 1 H), 7.36-7.29 (m, 1 H), 7.25 7.18 (m, 2 H), 7.10 (t, 1 H), 6.84 (t, 1 H) , 5.89 (s, 2 H), 5.09 (br. s., 1 H), 4.37 (br. s., 1 H), 3.10 (t, 1 H), 2.74 (br. s., 1 H), 1.84-1.72 (m, 3 H), 1.50 (br. s., 1 H), 1.19 (d, 3 H).

Reference compound I-312

The title compound was prepared following general procedure B, except that 3-azabicyclo[3.1.0]hexane-1-carboxylic acid (as the HCl salt) was the amine reagent, the contents were heated to 100 °C for 18 h and the aqueous layer during processing was treated with sodium chloride. The crude material was purified by silica gel chromatography with a gradient of 0-10% methanol/dichloromethane providing the desired compound, reference compound I-312 (44 mg, 70% yield) as an off-white solid.

1H-NMR (500 MHz, DMSO-de) 812.65 (br. s., 1 H), 9.08 (d, 1 H), 8.26 (d, 1 H), 7.53 (s, 1 H), 7.35-7.30 (m, 1 H), 7.26 (d, 1 H), 7.25-7.20 (m, 1 H), 7.10 (td, 1 H) ), 6.83-6.79 (m, 1 H), 5.91 (s, 2 H), 4.09-3.98 (m, 3 H), 3.81 (br. s., 1 H), 2.22-2.17 (m, 1 H), 1.51 (dd, 1 H), 0.97 (t, 1 H).

Reference compound I-313

The title compound was prepared following general procedure B, except that (S)-3-aminopropane-1,2-diol acid was the amine reagent, the content was heated at 100 °C for 20 h and the aqueous layer for processing was treated with sodium chloride. The crude material was purified by silica gel chromatography with a gradient of 0-10% methanol/dichloromethane providing the desired compound, reference compound I-313 (39 mg, 85% yield) as an off-white solid.1

1H-NMR (500 MHz, CD3OD) 88.74 (d, 1 H), 8.07 (d, 1 H), 7.42 (s, 1 H), 7.29-7.22 (m, 1 H), 7.11-7.05 (m, 1 H), 7.02 (td, 1 H), 6.88 (d, 1 H), 6.81 (td, 1 H), 5.95 (s, 2 H), 3.88 (quin, 1 H), 3.81-3.74 (m, 1 H), 3.69-3.62 (m, 1 H), 3.59 (s , 1H), 3.58 (s, 1H).

Reference compound I-314

The title compound was prepared following general procedure B, except that c/s-4-methylpyrrolidine-3-carboxylic acid was the amine reagent, the content was heated at 100 °C for 20 h, and the aqueous layer during processing It was treated with sodium chloride. The crude material was purified by silica gel chromatography with a gradient of 0-10% methanol/dichloromethane providing the desired compound, reference compound I-314 (50 mg, 72% yield) as an off-white solid.

1H-NMR (500 MHz, CD3OD) 68.74 (d, 1 H), 8.09 (d, 1 H), 7.41 (s, 1 H), 7.29-7.23 (m, 1 H), 7.11-7.06 (m, 1 H), 7.02 (t, 1 H), 6.91 (d, 1 H), 6.81 (t, 1 H), 5.95 (s, 2 H), 4.22-4.13 (m, 2 H), 3.98-3.92 (m, 1 H), 3.41 (t, 1 H), 2.84-2 .77 (m, 1 H), 2.58 (d, 1 H), 1.24 (d, 3 H).

Reference compound I-315

The title compound was prepared following general procedure B, except that serinol was the amine reagent, the content was heated at 100 °C for 20 h, and the aqueous layer during processing was treated with sodium chloride. The crude material was purified by silica gel chromatography with a gradient of 0-10% methanol/dichloromethane providing the desired compound, reference compound I-315 (49 mg, 84% yield) as an off-white solid.

1H-NMR (500 MHz, CD3OD) 68.75 (t, 1 H), 8.08 (dd, 1 H), 7.46 (d, 1 H), 7.30-7.23 (m, 1 H), 7.09 (dd, 1 H), 7.03 (t, 1 H), 6.91-6.88 (m, 1 H), 6.80 (t, 1 H), 5.96 (s, 2 H), 4.54 (quin, 1 H), 3.75-3.82 (m, 4 H).

Reference compound I-316

The title compound was prepared following general procedure B, except that acid (R)-3-aminopropane-1,2-diol (2 equiv.) was the amine reagent, the contents were heated at 100 °C for 20 h and the aqueous layer during processing was treated with sodium chloride. The crude material was purified by silica gel chromatography with a gradient of 0-10% methanol/dichloromethane providing the desired compound, reference compound I-316 (36 mg, 78% yield) as an off-white solid.

1H-NMR (500 MHz, CD3OD) 68.73 (d, 1 H), 8.06 (d, 1 H), 7.41 (s, 1 H), 7.27-7.22 (m, 1 H), 7.10-7.04 (m, 1 H), 7.01 (t, 1 H), 6.86 (d, 1 H), 6.83-6.78 (m, 1 H) , 5.94 (s, 2 H), 3.88 (quin, 1 H), 3.80-3.74 (m, 1 H), 3.68-3.62 (m, 1 H), 3 .58 (d, 2H).

Reference compound I-317

The title compound was prepared following general procedure B, except that 4-(aminomethyl)-2,6-difluorophenol was the amine reagent, the content was heated at 100 °C for 20 h, and the aqueous layer during preparation was treated with sodium chloride. The crude material was purified by silica gel chromatography with a gradient of 0-30% (acetonitrile:methanol=7:1)/dichloromethane providing the desired compound, the reference compound I-317 (38 mg, 30% yield) as a whitish solid.

1H-NMR (500 MHz, CD3OD) 68.77-8.74 (m, 1 H), 8.08 (d, 1 H), 7.37 (s, 1 H), 7.26 (dd, 1 H), 7.11-7.06 (m, 1 H), 7.06 7.01 (m, 3 H), 6.88 (d, 1 H), 6.84 (t, 1 H), 5.96 (s, 2 H), 4.69 (s, 2 H).

Reference compound I-318

The title compound was prepared following general procedure B, except that c/s-piperidin-2,4-diyldimethanol was the amine reagent and the contents were heated at 100 °C for 20 h. The reaction was poured into a 1:1 mixture of dichloromethane and water for processing, and the aqueous layer was treated with sodium chloride before extraction with dichloromethane. The crude material was purified by silica gel chromatography with a gradient of 0-70% (acetonitrile:methanol=7:1)/dichloromethane providing the desired compound, reference compound I-318 (39 mg, 25% yield). as a whitish solid. 1H-NMR (500 MHz, CD3OD) 68.75 (d, 1 H), 8.12 (d, 1 H), 7.40 (s, 1 H), 7.29-7.23 (m, 1 H), 7.11-7.05 (m, 1 H), 7.04-6.99 (m, 1 H), 6.90 (d, 1 H), 6.82 (td, 1 H) , 5.99-5.91 (m, 2 H), 4.52-4.45 (m, 1 H), 4.35-4.26 (m, 1 H), 3.86-3.76 (m, 2 H), 3.58-3.42 (m, 3 H), 2.09-1.99 (m, 2 H), 1.85-1.75 (m, 1 H), 1 .65-1.55 (m, 1 H), 1.45-1.36 (m, 1 H).

Reference compound I-319

The title compound was prepared following general procedure B, except that 3-phenylpiperidine-2-carboxylic acid (as the AcOH salt) was the amine reagent, the contents were heated at 100 °C for 20 h and the aqueous layer During preparation it was treated with sodium chloride. A portion of the crude material was purified by reverse phase HPLC using a gradient of 5-75% acetonitrile/water providing the desired compound, reference compound I-319 (30 mg, 9% yield) as an off-white solid.

1H-NMR (500 MHz, CDCh) 68.37 (d, 1 H), 8.27 (d, 1 H), 7.34-7.28 (m, 4 H), 7.26-7.22 (m, 1 H), 7.20 (s, 1 H), 7.15 (ddd, 1 H), 6.99-6.88 (m, 3 H), 6.45 (d, 1 H) , 5.91-5.82 (m, 2 H), 5.18 (d, 1 H), 4.31 (d, 1 H), 3.59 (td, 1 H), 3.26-3 .17 (m, 1 H), 2.49 (qd, 1 H), 2.06-1.99 (m, 1 H), 1.98-1.81 (m, 2 H).

Reference compound I-320

The title compound was prepared following general procedure B, except that (S)-3-(methylamino)p clothes -1,2 -diol was the amine reagent and the contents were heated at 100 °C for 20 h. The reaction was poured into a 1:1 mixture of dichloromethane and water for processing, and the aqueous layer was treated with sodium chloride before extraction with dichloromethane. The crude material was purified by silica gel chromatography with a gradient of 0-10% methanol/dichloromethane providing the desired compound, reference compound I-320 (81 mg, 84% yield) as a white solid.

1H-NMR (500 MHz, CD3OD) 88.75 (d, 1 H), 8.10 (d, 1 H), 7.42 (s, 1 H), 7.29-7.23 (m, 1 H), 7.11-7.06 (m, 1 H), 7.02 (t, 1 H), 6.88 (d, 1 H), 6.85-6.80 (m, 1 H) , 5.95 (s, 2 H), 4.03-3.94 (m, 2 H), 3.73-3.66 (m, 1 H), 3.58 (d, 2 H), 3 .42 (d, 3H).

Reference compound I-321

The title compound was prepared following general procedure B, except that (R)-3-(methylamino)propane-1,2-diol was the amine reagent and the contents were heated at 100 °C for 2 d. The reaction was poured into a 1:1 mixture of dichloromethane and water for processing, and the aqueous layer was treated with sodium chloride before extraction with dichloromethane. The crude material was purified by silica gel chromatography with a gradient of 0-10% methanol/dichloromethane providing the desired compound, reference compound I-321 (88 mg, 77% yield) as a white solid.

1H-NMR (500 MHz, CD3OD) 88.74 (d, 1 H), 8.09 (d, 1 H), 7.41 (s, 1 H), 7.28-7.22 (m, 1 H), 7.10-7.05 (m, 1 H), 7.04 6.99 (m, 1 H), 6.87 (d, 1 H), 6.82 (td, 1 H), 5.94 (s, 2 H), 4.02-3.93 (m, 2 H), 3.72-3.66 (m, 1 H), 3.58 (d, 2 H), 3, 41 (d, 3 H).

Reference compound I-322

The title compound was prepared following general procedure B, except that (S)-3-((cyclopropylmethyl)amino)propane-1,2-diol was the amine reagent and the contents were heated at 100 °C for 20 h. The reaction was poured into a 1:1 mixture of dichloromethane and water for processing, and the aqueous layer was treated with sodium chloride before extraction with dichloromethane. The crude material was purified by silica gel chromatography with a gradient of 0-10% methanol/dichloromethane providing the desired compound, reference compound I-322 (39 mg, 62% yield) as a white foam.

1H-NMR (500 MHz, CDCla) 88.45 (d, 1 H), 8.12 (d, 1 H), 7.29 (s, 1 H), 7.24-7.19 (m, 1 H), 7.05-6.97 (m, 3 H), 6.57 (d, 1 H), 5.99-5.94 (m, 1 H), 5.91-5.86 (m , 1 H), 4.14 (dd, 1 H), 4.02 (br. s., 1 H), 3.93 (br. s., 1 H), 3.86 (br. s., 1 H), 3.68-3.57 (m, 4 H), 3.43 (ddd, 1 H), 1.15-1.06 (m, 1 H), 0.65-0.53 ( m, 2 H), 0.38-0.32 (m, 1 H), 0.32-0.26 (m, 1 H).

Reference compound I-323

The title compound was prepared following general procedure B, except that (S)-3-(isopropylamino)propane-1,2-diol was the amine reagent and the contents were heated at 100 °C for 20 h. The reaction was poured into a 1:1 mixture of dichloromethane and water for processing, and the aqueous layer was treated with sodium chloride before extraction with dichloromethane. The crude material was purified by silica gel chromatography with a gradient of 0-50% (acetonitrile:methanol=7:1)/dichloromethane providing the desired compound, reference compound I-323 (12 mg, 20% yield). like a white solid.

1H-NMR (500 MHz, CDCh) 88.45 (d, 1 H), 8.13 (d, 1 H), 7.30 (s, 1 H), 7.25-7.19 (m, 1 H), 7.06-6.98 (m, 3 H), 6.58 (d, 1 H), 6.00-5.94 (d, 1 H), 5.91-5.85 (d , 1 H), 4.95 (br. s., 1 H), 4.67-4.58 (m, 1 H), 3.82-3.74 (m, 2 H), 3.70- 3.60 (m, 2 H), 3.59-3.49 (m, 2 H), 1.32 (d, 3 H), 1.29 (d, 3 H).

Reference compound I-324

The title compound was prepared following general procedure B, except that 2-(aminomethyl)-1,1,1,3,3,3-hexafluoropropan-2-ol was the amine reagent, the content was heated to 100 °C for 20 h and the aqueous layer was treated with sodium chloride before extraction with dichloromethane during processing. The crude material was purified by silica gel chromatography with a gradient of 0-10% ethyl acetate/hexane providing the desired compound, reference compound I-324 (5 mg, 9% yield) as a white solid.12

1H-NMR (500 MHz, CDCh) 88.48 (d, 1 H), 8.36 (s, 1 H), 8.26 (d, 1 H), 7.25-7.20 (m, 2 H), 7.14 (t, 1 H), 7.05-6.99 (m, 2 H), 6.59 (d, 1 H), 5.94 (s, 2 H), 5.59 (br. s., 1 H), 4.12 (d, 2 H).

Reference compound I-325

The title compound was prepared following general procedure B, except that 1-amino-2-methylpropan-2-ol was the amine reagent and the contents were heated at 100 °C for 20 h. The reaction was poured into a 1:1 mixture of dichloromethane and water for processing, and the aqueous layer was treated with sodium chloride before extraction with dichloromethane. The crude material was purified by silica gel chromatography with a gradient of 0-10% methanol/dichloromethane providing the desired compound, reference compound I-325 (43 mg, 93% yield) as a white solid.

1H-NMR (500 MHz, CDCI3) 58.43 (d, 1 H), 8.14 (d, 1 H), 7.27 (s, 1 H), 7.21-7.15 (m, 1 H), 7.04-6.98 (m, 1 H), 6.95 (t, 1 H), 6.84 (t, 1 H), 6.59 (d, 1 H), 5.95 (s, 2 H), 5.62 (br. s., 1 H), 3.70 (s, 1 H), 3.63 (d, 2 H), 1.31 (s, 6 H).

Reference compound I-326

A mixture of (S)-trifluorolactic acid (1.5 equiv.) and 1,1'-carbodiimidazole (1.5 equiv.) in THF was heated at 70 °C for 2 h. 2-(1-(2-fluorobenzyl)-5-(isoxazol-3-yl)-1H-pyrazol-3-yl)-pyrimidin-4 amine (intermediate product described in WO2012/3405 A1) (1 equiv.) added to the reaction mixture, and the contents were stirred at 70 °C for 3 d. The contents were cooled to 23 °C, diluted with ethyl acetate and washed with 1N HCl solution. The organic layer was washed with brine, dried over MgSO4, filtered and concentrated in vacuo. The crude material was purified by silica gel chromatography with a gradient of 0-100% ethyl acetate/hexanes providing the desired compound, reference compound I-326 (3 mg, 4% yield) as a white solid. 1H-NMR (500 MHz, CDCh) 5 9.61 (br. s., 1 H), 8.76 (d, 1 H), 8.49 (d, 1 H), 8.16 (d, 1 H), 7.47 (s, 1 H), 7.25-7.21 (m, 1 H), 7.11-7.03 (m, 1 H), 7.00 (t, 1 H) , 6.83 (t, 1 H), 6.63 (d, 1 H), 6.36 (br. s., 1 H), 6.06-5.95 (m, 2 H), 4, 69 (d, 1H).

Reference compound I-329 and reference compound I-330

Reference compound I-161 was dissolved by chiral separation with chiralcel-ODH 20 mm x 250 mm semipreparative column using a gradient of 10-90% isopropanol/hexanes. Collection of the first eluting peak and concentration in vacuo produced the reference compound I-329 as a white solid. Collection of the second eluting peak and concentration in vacuo yielded compound 330 as a white solid.

1H-NMR for Reference Compound I-329 (500 MHz, CDCh) 58.38 (d, 1 H), 8.16 (d, 1 H), 7.21 (s, 1 H), 7.20 7.15 (m, 1 H), 7.04-6.99 (m, 1 H), 6.97-6.93 (m, 1 H), 6.89-6.84 (m, 1 H) ), 6.48 (d, 1 H), 6.06-6.00 (m, 1 H), 5.93-5.88 (m, 1 H), 4.76 (d, 1 H), 4.15 (d, 1 H), 3.54-3.43 (m, 1 H), 2.09-1.98 (m, 1 H), 1.91-1.82 (m, 1 H) ), 1.81-1.64 (m, 3 H), 1.17 (d, 3 H).

1H-NMR for Reference Compound I-330 (500 MHz, CD3OD) 58.78-8.74 (d, 1 H), 8.21 (d, 1 H), 7.45 (s, 1 H) , 7.30-7.23 (m, 1 H), 7.11-7.06 (m, 1 H), 7.03 (t, 1 H), 6.86 (d, 1 H), 6 .83 (t, 1 H), 5.95 (s, 2 H), 5.04 (d, 1 H), 4.37 (d, 1 H), 3.69 (td, 1 H), 2 .16-2.07 (m, 1 H), 1.93-1.86 (m, 1 H), 1.82-1.70 (m, 2 H), 1.52 (qd, 1 H) , 1.21 (d, 3H).

Reference compound I-331 and reference compound I-332

A mixture of reference compound I-329 and 1,1-carbodiimidazole (1 equiv.) in DCM was heated to 45 °C until all starting material was consumed as observed by LC/MS. Cyclopropanesulfonamide (4 equiv.) and DBU (2 equiv.) were added to the reaction mixture, and the contents were stirred at 45 °C for an additional 30 min. The contents were cooled to 23 °C, quenched with 1N HCl solution, and the layers were separated. The aqueous layer was extracted with dichloromethane (2 times) and the organic parts were combined and washed with brine. The mixture was dried over Na2SO4, filtered and concentrated in vacuo. The crude material was purified by silica gel chromatography with a gradient of 0-20% (acetonitrile:methanol=7:1)/dichloromethane providing the reference compound I-331 (40 mg, 13% yield) as a white solid. , and the reference compound I-332 (3 mg, 1% yield) as a white solid.

1H-NMR for Reference Compound I-331 (400 MHz, DMSO-d6) 5 12.09 (s, 1 H), 9.14 (d, 1 H), 8.38 (d, 1 H), 7.25-7.18 (m, 1 H), 7.37-7.29 (m, 1 H), 7.55 (s, 1 H), 7.12-7.06 (m, 2 H ), 6.87-6.80 (m, 1 H), 5.94-5.84 (m, 2 H), 4.73 (d, 1 H), 4.23 (br. s., 1 H), 3.66-3.54 (m, 1 H), 2.94-2.85 (m, 1 H), 2.38 (d, 1 H), 1.89-1.81 (m , 1 H), 1.68-1.51 (m, 3 H), 1.15 (d, 3 H), 0.97-0.92 (m, 2 H), 0.89-0.84 (m, 2H).

1H NMR for reference compound I-332 (400 MHz, DMSO-d6) 511.97 (s, 1 H), 9.14 (d, 1 H), 8.41 (d, 1 H), 7. 65 (s, 1 H), 7.37-7.28 (m, 1 H), 7.26-7.17 (m, 1 H), 7.14-7.05 (m, 2 H), 6.88-6.79 (m, 1 H), 5.97-5.86 (m, 2 H), 4.70 (d, 1 H), 4.13 (d, 1 H), 3, 83-3.72 (m, 1 H), 2.97-2.87 (m, 1 H), 2.10 (d, 1 H), 1.85 (d, 1 H), 1.74- 1.60 (m, 2 H), 1.53-1.39 (m, 1 H), 1.16 (d, 3 H), 1.05-0.93 (m, 3 H), 0, 87-0.78 (m, 1H).

Reference compound I-333

A mixture of 3-(3-(4-chloropyrimidin-2-yl)-1-(2-fluorobenzyl)-1H-pyrazol-5-yl)isoxazole (the synthesis of which is described in the procedure for the reference compound I- 24) (1 equiv.), (2R,3S)-3-methylpiperidine-2-carboxylic acid (as (1S)-(+)-camphorsulfonic acid salt, 1 equiv.), and triethylamine (1 equiv.) was heated at 110 °C for 48 h as a solution in dioxane/water (2:1). The contents were cooled to 23 °C, and divided between a 1:1 mixture of dichloromethane and 1N HCl solution. The layers were separated and the aqueous layer was extracted with dichloromethane (2 times), and the organic parts were combined and washed with brine. The mixture was dried over Na2SO4, filtered and concentrated in vacuo. The crude material was purified by silica gel chromatography with a gradient of 0-20% (acetonitrile:methanol=7:1)/dichloromethane providing the desired compound, reference compound I-333 (10 mg, 4% yield). like a white solid.

1H-NMR (400 MHz, DMSO-d6) 512.64 (br. s., 1 H), 9.10 (d, 1 H), 8.30 (d, 1 H), 7.55 (br. s., 1 H), 7.38-7.29 (m, 1 H), 7.27-7.16 (m, 2 H), 7.15-7.06 (m, 1 H), 6 .83 (d, 2 H), 5.90 (s, 2 H), 3.21 (s, 1 H), 1.94 (br. s., 1 H), 1.89-1.80 ( m, 1 H), 1.69-1.35 (m, 5 H), 1.13 (d, 3 H).

I-334 REFERENCE COMPOSITION

The title compound was prepared following general procedure B, except that 2-amino-2-(hydroxymethyl)propane-1,3-diol was the amine reagent and the contents were heated at 110 °C for 20 h. The crude material was purified by silica gel chromatography using a gradient of 0-20% (acetonitrile:methanol=7:1)/dichloromethane providing the desired compound, the desired reference compound I-334 (185 mg, 72% yield). ) as a whitish solid.

1H-NMR (400 MHz, DMSO-da) 89.10 (d, 1 H), 8.24 (d, 1 H), 7.46 (s, 1 H), 7.36-7.29 (m , 1 H), 7.25-7.18 (m, 2 H), 7.10 (td, 1 H), 6.91-6.84 (m, 1 H), 6.36 (s, 1 H), 5.86 (s, 2 H), 4.95 (br. s., 3 H), 3.76-3.72 (m, 6 H).

Reference compound I-335

The title compound was prepared following general procedure B, except that (S)-2-amino-3-hydroxypropanamide was the amine reagent and the contents were heated at 110 °C for 20 h. The crude material was purified by silica gel chromatography using a gradient of 0-20% (acetonitrile:methanol=7:1)/dichloromethane providing the desired compound, reference compound I-335 (170 mg, 37% yield). like a white solid.

1H-NMR (400 MHz, DMSO-d6) 89.14 (d, 1 H), 8.56 (br. s., 1 H), 8.41 (d, 1 H), 7.70 (s, 2 H), 7.37-7.31 (m, 1 H), 7.29 7.19 (m, 3 H), 7.14-7.08 (m, 1 H), 6.86 (t , 1 H), 5.94 (s, 2 H), 4.87-4.80 (m, 1 H), 3.89-3.78 (m, 2 H), 3.06 (qd, 1 H).

Reference compound I-336

A solution of the reference compound I-112 (1 equiv.) in DMF was successively treated with Hunig's base (3 equiv.) and HATU (1 equiv.). After stirring for 5 min, serinol (1.5 equiv.) was added, and the reaction was stirred at 23 °C for 20 h. The mixture was partitioned between a 1:1 mixture of dichloromethane and a 1N HCl solution. The layers were separated and the aqueous layer was extracted with dichloromethane (2 times). The combined organic portions were washed with brine. The mixture was dried over Na2SO4, filtered and concentrated in vacuo. The crude material was purified by silica gel chromatography with a gradient of 0-20% (acetonitrile:methanol=7:1)/dichloromethane providing the desired compound, reference compound I-336 (22 mg, 26% yield). like a white solid.

1H-NMR (400 MHz, DMSO-d6) 89.14 (d, 1 H), 8.96 (br. s., 1 H), 8.45 (d, 1 H), 8.17 (d, 1 H), 7.85 (s, 1 H), 7.38-7.29 (m, 1 H), 7.28-7.18 (m, 2 H), 7.14-7.07 ( m, 1 H), 6.86 (t, 1 H), 5.96 (s, 2 H), 4.94-4.86 (m, 1 H), 3.86-3.78 (m, 2 H), 3.74-3.65 (m, 1 H), 3.64-3.53 (m, 1 H), 3.53-3.46 (m, 1 H), 3.45- 3.40 (m, 2 H), 3.40-3.34 (m, 2 H), 3.16-3.04 (m, 1 H).

Reference compound I-337

The title compound was prepared following general procedure B, except that 3-amino-1,1,1-trifluoropropan-2-ol (5 equiv.) was the amine reagent and the contents were heated to 110 °C for 8 p.m. The crude material was purified by silica gel chromatography using a gradient of 0-20% (acetonitrile:methanol=7:1)/dichloromethane providing the desired compound, reference compound I-337 (70 mg, 53% yield). like a white solid.

1H-NMR (400 MHz, CDCla) 88.76 (d, 1 H), 8.10 (d, 1 H), 7.38 (s, 1 H), 7.30-7.23 (m, 1 H), 7.08 (dd, 1 H), 7.05-7.00 (m, 1 H), 6.88-6.82 (m, 2 H), 5.95 (s, 2 H) , 4.40-4.30 (m, 1 H), 3.99 (dd, 1 H), 3.70 (dd, 1 H).

Reference compound I-339

The title compound was prepared in 2 steps:

Step 1: Synthesis of 2-(5-fluoro-2-(1-(2-fluorobenzyl)-5-(isoxazol-3-yl)-1H-pyrazol-3-yl)pyrimidin-4-yl)-1, methyl 2,3,4-tetrahydroisoquinoline-8-carboxylate

This intermediate was prepared following general procedure B, except that methyl 1,2,3,4-tetrahydroisoquinoline-8-carboxylate (as the HCl salt) was the amine reagent. The preparation gave the desired methyl ester, 2-(5-fluoro-2-(1-(2-fluorobenzyl)-5-(isoxazol-3-yl)-1H-pyrazol-3-yl)pyrimidin-4-yl) Methyl -1,2,3,4-tetrahydroisoquinoline-8-carboxylate (reference compound I-338, 55 mg, yield 97%) as an orange oil, and was collected separately without further purification.

Stage 2: Synthesis of reference compound I-339

A solution of 2-(5-fluoro-2-(1-(2-fluorobenzyl)-5-(isoxazol-3-yl)-1H-pyrazol-3-yl)pyrimidin-4-yl)-1,2, Methyl 3,4-tetrahydroisoquinoline-8-carboxylate and hydrated lithium hydroxide (1.5 equiv.) in tetrahydrofuran, water and methanol (3:1:1 ratio) were stirred at 23 °C for 21 h. More base (1.5 equiv.) was added, and the solution was stirred for 24 h. The solution was poured into water, 1 N sodium hydroxide and dichloromethane (10:1:10 ratio). The layers were separated and the aqueous layer was acidified to pH 1. The aqueous layer was extracted with dichloromethane, dried with magnesium sulfate, filtered, and the solvent was removed in vacuo providing the desired compound, reference compound I- 339 (9 mg, 17% yield in 2 steps) as a white solid.

1H-NMR (400 MHz, DMSO-de) 813.14 (s, 1 H), 9.12 (d, 1 H), 8.32 (d, 1 H), 7.79-7.77 (m , 1 H), 7.48 (s, 1 H), 7.43-7.41 (d, 1 H), 7.34-7.30 (m, 2 H), 7.24-7.20 (m, 2 H), 7.11 (dt, 1 H), 6.85 (dt, 1 H), 5.89 (s, 2 H), 5.34 (d, 2 H), 4.07 (t, 2H), 3.04 (t, 2H).

Reference compound I-341

The title compound was prepared in 3 steps:

Stage 1: Synthesis of c/s-1-(5-fluoro-2-(1-(2-fluorobenzyl)-5-(isoxazol-3-yl)-1H-pyrazol-3-yl)pyrimidin-4yl)- 3-methylpiperidine-2-carboxamide

(Reference compound I-340)

To a solution of the reference compound I-161 and triethylamine (1 equiv.) at 0 °C in tetrahydrofuran, ethyl chloroformate (1.05 equiv.) was added dropwise over 5 minutes. The reaction mixture was kept at 0 °C for 45 min, and then ammonium hydroxide (7 equiv.) was added. The solution was immediately heated to 23°C and stirred for an additional 15 hours. The reaction mixture was diluted with ethyl acetate and saturated aqueous ammonium chloride. The layers were separated and the aqueous layer was extracted with ethyl acetate. The organic substances were dried over magnesium sulfate, filtered, and the solvent was removed in vacuo. Purification by silica gel chromatography (20-100% hexanes gradient in ethyl acetate) afforded c/s-1-(5-fluoro-2-(1-(2-fluorobenzyl)-5-(isoxazole-3 -yl)-1H-pyrazol-3-yl)pyrimidin-4-yl)-3-methylpiperidine-2-carboxamide (reference compound I-340, 270 mg, yield 54%) as a yellow foam.

1H NMR (400 MHz, CDCla) 8 ppm 8.46 (d, 1 H), 8.17 (d, 1 H), 7.23 (s, 1 H), 7.22-7.17 (m, 1 H), 7.12-6.91 (m, 3 H), 6.59 (d, 1 H), 6.06-5.76 (m, 2 H), 5.34 (br. s. , 1 H), 4.76 (d, 1 H), 4.18 (d, 1 H), 3.32 (ddd, 1 H), 2.64-2.50 (m, 1 H), 2 .08-1.79 (m, 2 H), 1.65-1.50 (m, 1 H), 1.49-1.37 (m, 1 H), 1.15 (d, 1 H) , 1.05 (d, 3H).

Step 2: Synthesis of c/s-1-(5-fluoro-2-(1-(2-fluorobenzyl)-5-(isoxazol-3-yl)-1H-pyrazol-3-yl)pyrimidin-4-yl )-3-methylpiperidin-2-carbonitrile

To a 0 °C solution of c/s-1 -(5-fluoro-2-(1 -(2-fluorobenzyl)-5-(isoxazol-3-yl)-1H-pyrazol-3-yl)pyrimidin- 4-yl)-3-methylpiperidine-2-carboxamide in pyridine trifluoroacetic anhydride (2 equiv.) was added dropwise over 5 minutes.

After stirring for 45 minutes at 0°C, the solution was warmed to room temperature and then immediately poured into dichloromethane and saturated aqueous ammonium chloride. The layers were separated and the aqueous layer was extracted with dichloromethane. The organic substances were dried over magnesium sulfate, filtered, and the solvent was removed in vacuo. Purification by silica gel chromatography (10-75% hexanes gradient in ethyl acetate) afforded c/s-1 -(5-fluoro-2-(1 -(2-fluorobenzyl)-5-(isoxazole-3 -yl)-1H-pyrazol-3-yl)pyrimidin-4-yl)-3-methylpiperidin-2-carbonitrile (215 mg, yield 83%) as a white foam.

1H NMR (400 MHz, CDCla) 8 ppm 8.44 (d, 1 H), 8.31 (d, 1 H), 7.30 (s, 1 H), 7.22-7.11 (m, 1 H), 7.06-6.91 (m, 2 H), 6.82 (t, 1 H), 6.59 (d, 1 H), 5.95 (s, 2 H), 5, 40 (s, 1 H), 4.35 (d, 1 H), 3.32 (td, 1 H), 2.48-2.35 (m, 1 H), 2.20-1.84 ( m, 2 H), 1.61-1.76 (m, 2 H), 1.17 (d, 3 H).

Stage 3: Synthesis of reference compound I-341

A suspension of c/s-1-(5-fluoro-2-(1-(2-fluorobenzyl)-5-(isoxazol-3-yl)-1H-pyrazol-3-yl)pyrimidin-4-yl)- 3-methylpiperidin-2-carbonitrile, ammonium hydrochloride (5 equiv.), and sodium azide (5 equiv.) in W,A/-dimethylformamide was heated at 90 °C for 60 hours. The solution was diluted with ethyl acetate and 1N aqueous hydrochloric acid solution. The layers were separated and the aqueous layer was extracted with ethyl acetate. The organic substances were washed with water and brine, dried over magnesium sulfate, filtered, and the solvent was removed in vacuo. The crude solid was suspended in dichloromethane and filtered to give the by-product contaminated product. The filtrate was concentrated in vacuo and the resulting solid was suspended in diethyl ether and filtered to provide the desired compound, reference compound I-341 (135 mg, 57% yield) as a white solid.

1H-NMR (400 MHz, CDCh) 88.51 (d, 1 H), 8.15 (d, 1 H), 7.29 (s, 1 H), 7.25-7.20 (m, 2 H), 7.04-6.99 (m, 2 H), 6.63 (d, 1 H), 6.09 (d, 1 H), 6.00 (d, 1 H), 5.18 (d, 1 H), 3.98 (dd, 1 H), 3.31 (dt, 1 H), 2.81-2.75 (m, 1 H), 2.54-2.46 (m , 1 H), 2.13-2.02 (m, 1 H), 1.80-1.75 (m, 1 H), 1.60-1.51 (m, 1 H), 1.11 (d, 3H).

Reference compound I-342

The title compound was prepared following general procedure B, except that 1-((methylamino)methyl)cyclobutanecarboxylic acid (as the TFA salt) was the amine reagent. The preparation provided the desired compound, reference compound I-342 (64 mg, quantitative yield) as a yellow solid.

1 H - R M N ( 400 M H z , C D 3O D ) 8 8 , 81 ( d , 1 H ) , 8 , 29 ( d , 1 H ) , 7 , 56 ( m , 1 H ) , 7 , 32 - 7 , 26 (m, 1 H), 7, 11 - 7, 0 2 (m, 2 H), 6, 96 6, 92 (m, 2 H), 5, 98 (s, 2 H), 4, 37 ( s, 2H), 3, 48 (d, 3H), 2, 51 - 2, 44 (m, 2H), 2, 25 - 2, 16 (m, 2H), 2, 09 - 1, 93 (m, 2H).

REFERENCE COMPOSITION I-343

A suspension of 5-fluoro-2-(1-(2-fluorobenzyl)-5-(thiazol-2-yl)-1H-pyrazol-3-yl)pyrimidin-4-ol (intermediate previously described in WO2013/101830 A1) in phosphoryl chloride (20 equiv.) was heated at 60 °C for 2 h. The phosphoryl chloride was removed under a stream of nitrogen and the resulting residue was dissolved in dioxane and water (2:1 ratio). After addition of 1-((methylamino)methyl)cyclopropanecarboxylic acid hydrochloride (3 equiv.) and triethylamine (10 equiv.), the solution was heated at 90 °C for 4 h. The solution was diluted with dichloromethane and 1 N hydrochloric acid solution. The layers were separated and the aqueous layer was extracted with dichloromethane. The organic substances were dried over magnesium sulfate, filtered, and the solvent was removed in vacuo. Purification by silica gel chromatography (0-5% methanol in dichloromethane gradient) provided the desired compound, reference compound I-343 (17 mg, 52% yield) as a white solid.

1H-NMR (400 MHz, DMSO-da) 812.34 (s, 1 H), 8.23 (d, 1 H), 7.98 (d, 1 H), 7.92 (d, 1 H) , 7.35 (s, 1 H), 7.34-7.29 (m, 1 H), 7.23-7.18 (m, 1 H), 7.10 (dt, 1 H), 6 .93 (dt, 1 H), 6.02 (s, 2 H), 4.00 (s, 2 H), 3.24 (d, 3 H), 1.15-1.12 (m, 2 H), 1.03-1.01 (m, 2 H).

Reference compound I-344

To a solution of the reference compound I-248 in dichloromethane was added carbonyldiimidazole (1.2 equiv.). The resulting mixture was stirred at 40 °C for 45 min. The solution was cooled to 23 °C and 1,8-diazabicyclo[5.4.0]undec-7-ene (2 equiv.) was added, followed by cyclopropanesulfonamide (3 equiv.). After stirring for 16 h, the solution was poured into dichloromethane and 1 N hydrochloric acid solution. The layers were separated and the aqueous layer was extracted with dichloromethane. The organic substances were dried over magnesium sulfate, filtered, and the solvent was removed in vacuo. Initial purification by silica gel chromatography (methanol in dichloromethane) gave the impure product. The impure residue was taken up in diethyl ether, and hexanes were added until the solution became cloudy. After stirring for 20 minutes, the solid was filtered to provide the desired compound, reference compound I-344 (29 mg, 48% yield) as a white solid.

1H-NMR (400 MHz, CDCla) 88.44 (d, 1 H), 8.09 (d, 1 H), 7.23-7.17 (m, 2 H), 7.03-6.97 (m, 3 H), 6.51 (d, 1 H), 5.95 (s, 2 H), 3.96 (dd, 1 H), 3.69-3.63 (m, 1 H) , 3.32 (d, 3 H), 2.90-2.81 (m, 2 H), 2.03-1.96 (m, 1 H), 1.18-1.14 (m, 2 H), 1.03 (dd, 6 H), 0.92-0.90 (m, 2 H).

Reference compound I-345

A suspension of 5-fluoro-2-(1-(2-fluorobenzyl)-5-(thiazol-4-yl)-1H-pyrazol-3-yl)pyrimidin-4-ol (intermediate previously described in WO2013/101830 A1) in phosphoryl chloride (50 equiv.) was heated at 60 °C for 2 h. The phosphoryl chloride was removed under a stream of nitrogen, and the resulting residue was dissolved in dioxane and water (2:1) and treated with 1-((methylamino)methyl)cyclopropanecarboxylic acid hydrochloride (3 equiv.), followed by of triethylamine (10 equiv.). The resulting solution was heated at 90 °C for 7.5 days. The reaction mixture was poured into dichloromethane and 1N hydrochloric acid solution. The layers were separated and the aqueous layer was extracted with dichloromethane. The organic substances were dried over magnesium sulfate, filtered, and the solvent was removed in vacuo. Purification by silica gel chromatography (0-10% methanol in dichloromethane gradient) provided the reference compound I-345 (2.8 mg, 11% yield) as a white film.

1H-MRI (400 MHz, CD3OD) 89.03 (d, 1 H), 8.07 (d, 1 H), 7.89 (d, 1 H), 7.24-7.19 (m, 2 H), 7.04-6.96 (m, 2 H), 6.81 (t, 1 H), 5.94 (s, 2 H), 4.13 (s, 2 H), 3.34 (d, 3 H), 1.28-1.25 (m, 2 H), 1.08-1.05 (m, 2 H).

Reference compound I-346

The title compound was prepared following general procedure B, except that 3-(trifluoromethyl)-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridine (as the HCl salt) was the amine reagent. After preparation, the crude solid was suspended in diethyl ether and filtered to provide the desired compound, reference compound I-346 (57 mg, 84% yield) as a tan solid.

1H-NMR (400 MHz, CD3OD) 88.84 (d, 1 H), 8.41 (d, 1 H), 7.63 (s, 1 H), 7.33-7.28 (m, 1 H), 7.13-7.04 (m, 2 H), 6.98 6.95 (m, 2 H), 6.03 (s, 2 H), 5.19 (s, 2 H), 4.42 (t, 2H), 3.09 (t, 2H).

Reference compound I-347

The title compound was prepared following general procedure B, except that syn-piperidine-3,4,5-triol was the amine reagent. Upon completion of the reaction, as determined by LC/MS, the solvent was removed in vacuo. Methanol was added and the resulting suspension was filtered to provide the desired compound, reference compound I-347 (20 mg, 11% yield) as a solid.

1 H -R M N (400 M H z, D M S O -d 6) 8 9 .09 (d, 1 H), 8 .28 (d, 1 H), 7 .52 (s, 1 H), 7 .35 -7 .29 (m, 1 H), 7 .24-7 .21 (m, 2 H), 7 .09 (t, 1 H), 6 .80 (t, 1 H), 5 .90 (s, 2 H), 4 .92 (d, 2 H), 4 .80 (d, 1 H), 4 .15-4 .11 (m, 2 H), 3 .83 -3 .80 (m, 1 H) , 3 .58 -3 .53 (m, 2 H), 3 .27-3 .21 (m, 2 H).

Reference compound I-348

The title compound was prepared following general procedure B, except that 1,3-diaminopropan-2-ol was the amine reagent and the contents were heated at 40 °C for 45 min. After completion of the reaction, as determined by LC/MS, the dioxane was removed in vacuo and enough methanol was added to solubilize the crude mixture. Purification by reverse phase HPLC (gradient of 5-75% acetonitrile in water with 0.1% trifluoacetic acid, 20 min) provided the desired compound, reference compound I-348 (58 mg, 80% yield) as a pink foam

1H-NMR (400 MHz, CD3OD) 88.82 (d, 1 H), 8.27 (d, 1 H), 7.62 (s, 1 H), 7.32-7.27 (m, 1 H), 7.12-7.03 (m, 2 H), 6.95 6.91 (m, 2 H), 6.02 (d, 1 H), 5.97 (d, 1 H), 4.21-4.15 (m, 1 H), 3.85-3.77 (m, 2 H), 3.22-3.18 (dd, 1 H), 2.96 (dd, 1 H ).

Reference compound I-349

To a solution of the reference compound I-340 and triethylamine (5 equiv.) in dichloromethane at 0 °C was added methanesulfonyl chloride (1 equiv.) dropwise. After stirring for 45 minutes, a saturated aqueous sodium bicarbonate solution was added and the reaction mixture was heated to 23 °C. Dichloromethane was added and the layers separated. The aqueous layer was extracted with dichloromethane and the organic substances were dried over magnesium sulfate, filtered, and the solvent was removed in vacuo. Purification by reverse phase HPLC (gradient of 5-50% acetonitrile in water with 0.1% TFA) provided the desired compound, reference compound I-349 (6 mg, 25% yield) as a white solid.

1H-NMR (400 MHz, CD3OD) 88.78 (d, 1 H), 8.24 (d, 1 H), 7.70 (s, 1 H), 7.31-7.25 (m, 1 H), 7.11-7.02 (m, 3 H), 6.92 6.88 (m, 1 H), 6.04 (s, 2 H), 4.10-4.01 (m, 2 H), 3.61-3.56 (m, 1 H), 3.18-3.17 (m, 2 H), 2.95 (s, 3 H).

Reference compound I-350

The title compound was prepared following general procedure B, except racemic cis-piperidin-3,4-diol (as the HCl salt) was the amine reagent. The crude material was purified by reverse phase HPLC (gradient of 5-50% acetonitrile in water with 0.1% TFA, 20 min) gave the desired compound, reference compound I-350 (1.7 mg, yield 3%) as a transparent film.

1H-NMR (400 MHz, CD3OD) 88.81 (m, 1 H), 8.28 (d, 1 H), 7.64 (s, 1 H), 7.32-7.26 (m, 1 H), 7.12-7.03 (m, 2 H), 6.97 6.93 (m, 2 H), 6.01 (s, 2 H), 4.52 (br s, 2 H) , 4.38 (dd, 1 H), 3.97-3.90 (m, 2 H), 3.77-3.72 (m, 1 H), 2.02-1.97 (m, 1 H), 1.90-1.86 (m, 1 H).

Reference compound I-351

The title compound was prepared following general procedure B, except that ferf-butyl 3-(aminomethyl)-3-hydroxyazetidine-1-carboxylate was the amine reagent, and the reaction was collected as a solution in dioxane. . Ethyl acetate was used as a solvent during preparation. The crude material was purified by silica gel chromatography (0-5% methanol in dichloromethane gradient) to provide the desired compound, reference compound I-351 (144 mg, quantitative yield) as a white solid.

1H-NMR (400 MHz, CD3OD) 88.76 (d, 1 H), 8.11 (d, 1 H), 7.44 (s, 1 H), 7.30-7.25 (m, 1 H), 7.12-7.01 (m, 2 H), 6.92 (d, 1 H), 6.81 (t, 1 H), 5.97 (s, 2 H), 4.09 (d, 2 H), 3.93 (br s, 2 H), 3.78 (d, 2 H), 1.40 (s, 9 H).

Reference compound I-352

The title compound was prepared following general procedure B, except that 3-(aminomethyl)oxetan-3-ol was the amine reagent, and the reaction was collected as a solution in dioxane. After stirring for 1.5 h at 90 °C, 2 more equivalents of oxetane were added and the reaction mixture was stirred at 90 °C for 3 h. Then, the reaction solution was poured into ethyl acetate and 1N aqueous hydrochloric acid solution. The layers were separated and the aqueous layer was extracted with 5:1 dichloromethane/isopropyl alcohol. The organic substances were dried over magnesium sulfate, filtered, and the solvent was removed in vacuo. Purification by reverse phase HPLC (gradient 5-75% acetonitrile in water with 0.1% TFA, 15 min) provided the reference compound I-352 (25 mg, 53% yield) as a white solid.1

1H-NMR (400 MHz, CD3OD) 88.83 (d, 1 H), 8.29 (d, 1 H), 7.65 (s, 1 H), 7.32-7.28 (m, 1 H), 7.13-7.05 (m, 2 H), 6.99 6.94 (m, 2 H), 6.02 (s, 2 H), 4.05 (d, 1 H), 3.98 (d, 1 H), 3.78-3.63 (m, 4 H).

Reference compound I-353

To a solution of the reference compound I-351 at 23 °C in dichloromethane was added trifluoacetic acid (30 equiv.) in one part. After stirring for 30 minutes, the solution was concentrated in vacuo and the resulting residue was taken up in diethyl ether. The solid was filtered to provide the desired compound, reference compound I-353 (160 mg, quantitative yield) as a tan solid.

1 H -R M N (400 M Hz, C D 3O D ) 8 8 .83 (m, 1 H), 8 .29 -8 .25 (m, 1 H), 7 .60 -7 .57 (m, 1 H ), 7 .33 -7 .28 (m , 1 H), 7 .14 -7 .04 (m, 2 H), 6 .93 -6 .89 (m , 2 H), 6 .00 (s, 2 H), 4.31 (d, 2 H), 4 .05 (m, 2 H), 3 .97 (d, 2 H).

REFERENCE COMPOSITION I-354

The title compound was prepared following general procedure B, except that N-(1-(aminomethyl)cyclopropyl)-1,1,1-trifluoromethanesulfonamide (as the HCl salt) was the amine reagent. The crude material was purified by silica gel chromatography (0-50% ethyl acetate in hexanes gradient) providing the desired compound, reference compound I-354 (15 mg, 25% yield) as a white solid.

1H-NMR (400 MHz, CD3OD) 88.75 (d, 1 H), 8.08 (d, 1 H), 7.50 (s, 1 H), 7.28-7.24 (m, 1 H), 7.10-7.07 (m, 1 H), 7.04 7.01 (m, 1 H), 6.83-6.80 (m, 2 H), 5.96 (s, 2 H), 3.85 (s, 2 H), 1.06-1.04 (m, 2 H), 0.99-0.96 (m, 2 H).

Reference compound I-355

The title compound was prepared following general procedure B, except that 2-(aminomethyl)-3,3,3-trifluoropropane-1,2-diol was the amine reagent, and the reaction was carried out as a solution in dioxane . After preparation with ethyl acetate and 1N hydrochloric acid solution, purification by silica gel chromatography (0-10% methanol in dichloromethane gradient) provided the desired compound, reference compound I-355 (17 mg, yield 36%) as a white solid.

1H-NMR (400 MHz, CD3OD) 88.75 (d, 1 H), 8.15 (d, 1 H), 7.39 (s, 1 H), 7.28-7.23 (m, 1 H), 7.09-7.01 (m, 2 H), 6.88 6.85 (m, 2 H), 5.96 (d, 1 H), 5.93 (d, 1 H), 4.02 (d, 1 H), 3.90 (d, 1 H), 3.75 (d, 1 H), 3.62 (d, 1 H).

Reference compound I-356

The title compound was prepared following general procedure B, except that (S)-3-aminopyrrolidin-2-one (as the HCl salt) was the amine reagent. The crude residue was suspended in 3:1 diethyl ether and dichloromethane. The resulting solid was filtered to provide the desired compound, reference compound I-356 (10 mg, 22% yield) as a tan solid.

1H-NMR (400 MHz, CD3OD) 88.83 (s, 1 H), 8.34 (m, 1 H), 7.63-7.62 (m, 1 H), 7.33-7.29 (m, 1 H), 7.13-7.05 (m, 2 H), 6.98-6.96 (m, 2 H), 6.03 (s, 2 H), 5.36-5 .32 (m, 1 H), 3.56-3.49 (m, 2 H), 2.66-2.61 (m, 1 H), 2.38-2.30 (m, 1 H) .

Reference compound I-357

The title compound was prepared in 5 steps:

Step 1: Synthesis of ferf-butyl 3-(2,22-trifluoro-1-hydroxyethyl)acetidine-1-carboxylate

To a 0 °C solution of ferf-butyl 3-formylazetidine-1-carboxylate (1 equiv.) and trimethyl(trifluoromethyl)silane (1.4 equiv.) in tetrahydrofuran was added detetrabutylammonium fluoride (1 M solution in tetrahydrofuran, 1.1 equiv.) over the course of 10 min. The solution was immediately heated to 23 °C and stirred for 19 h. The reaction mixture was poured into 1N hydrochloric acid and diethyl ether solution. The layers were separated and the aqueous layer was extracted with diethyl ether. The organic substances were washed with saturated aqueous sodium chloride, dried over magnesium sulfate, filtered, and the solvent was removed in vacuo to give 3-(2,2,2-trifluoro-1-hydroxyethyl)azetidine-1-carboxylate. of ferf-butyl (558 mg, 79% yield) as a pale yellow solid.

Step 2: Synthesis of fbutyl 3-(2,2,2-trifluoroacetyl)azetidine-1-carboxylate

To ferf-butyl 3-(2,2,2-trifluoro-1-hydroxyethyl)azetidine-1-carboxylate (1 equiv.) in dichloromethane at 0 °C was added Dess-Martin periodinane (2 equiv.) in a single part. After 5 minutes at 0 °C, the solution was warmed to 23 °C. After two hours, LC/MS analysis showed complete conversion. The reaction mixture was poured into a saturated aqueous solution of sodium dithionite and saturated aqueous sodium bicarbonate 5:1 (75 ml). After stirring for 10 minutes, dichloromethane was added and the layers separated. The aqueous layer was extracted with dichloromethane, the organic substances were dried over magnesium sulfate, filtered, and the solvent was removed in vacuo to provide ferf-butyl 3-(2,2,2-trifluoroacetyl)azetidine-1-carboxylate ( 263 mg, yield 92%) as a white greasy solid which was a mixture of ketone and hydrate according to 1H-NMR.

Step 3: Synthesis of ferf-butyl 3-(1,1,1,3,3,3-hexafluoro-2-hydroxypropan-2-yl)azetidine-1 carboxylate

To a 0 °C solution of ferf-butyl 3-(2,2,2-trifluoroacetyl)azetidine-1-carboxylate (1 equiv.) and trimethyl(trifluoromethyl)silane (1.4 equiv.) in tetrahydrofuran was added tetrabutylammonium fluoride (1.1 equiv.) as a solution in 1 M tetrahydrofuran dropwise over 5 min. The solution was immediately heated to 23 °C and stirred for 2.5 d. Then, the solution was poured into ethyl acetate and 1N aqueous hydrochloric acid solution. The layers were separated and the aqueous layer was extracted with ethyl acetate (2 times). The organic substances were washed with saturated aqueous sodium chloride, dried over magnesium sulfate, filtered, and the solvent was removed in vacuo. Purification by silica gel chromatography (gradient 0-60% ethyl acetate in hexanes) afforded 3-(1,1,1,3,3,3-hexafluoro-2-hydroxypropan-yl)azetidine-1-carboxylate of ferf-butyl (24 mg, 7% yield) as an oily solid.

Step 4: Synthesis of 2-(acetidin-3-yl)-1,1,1,3,3,3-hexafluoropropan-2-ol

Ferf-butyl 3-(1,1,1,3,3,3-hexafluoro-2-hydroxypropan-2-yl)azetidine-1-carboxylate in trifluoroacetic acid and dichloromethane (1:2 ratio) was stirred for 1. 5 hrs. The solution was then concentrated in vacuo to provide 2-(acetidin-3-yl)-1,1,1,3,3,3,3-hexafluoropropan-2ol as TFA salt.

Stage 5: Synthesis of reference compound I-357

The title compound was prepared following general procedure B, except that 2-(acetidin-3-yl)-1,1,1,3,3,3,3-hexafluoropropan-2ol (as the TFA salt, 1.5 equiv.) was the amine reagent. The crude residue was taken up in 1:1 dichloromethane and diethyl ether and the solid was filtered and washed with additional diethyl ether to provide the desired compound, reference compound I-357 (17 mg, 64% yield) as a white solid.

1H-NMR (400 MHz, CD3OD) 88.83 (d, 1 H), 8.27 (d, 1 H), 7.62 (s, 1 H), 7.35-7.30 (m, 1 H), 7.15-7.07 (m, 2 H), 7.00 6.96 (m, 2 H), 6.05 (s, 2 H), 4.82-4.70 (m, 4 H), 3.79 (quint, 1 H).

Reference compound I-359

This compound was prepared in two stages

Stage 1: I-358 ester synthesis

The title compound was prepared following general procedure B, except that 1-aminomethylcyclopropanol (2 equiv.) was the amine reagent, no triethylamine was used, and the contents were heated to 100 °C for 96 h as a solution in dioxane/water (3:1). Ethyl acetate was the solvent used for processing. The crude material was first purified by silica gel chromatography using a gradient of 0-100% ethyl acetate/hexanes, then by reverse phase HPLC (water/acetonitrile with 0.1% trifluoacetic acid). The water/acetonitrile fractions containing product were treated with 10% NaHCO3 (aq), concentrated to 5 ml, then extracted with ethyl acetate to recover the neutral material providing the desired compound, reference compound I-358 (32 mg, 28% yield) as a white solid.1

1H-NMR (400 MHz, CDCh) 88.42 (d, 1 H), 8.11 (m, 1 H), 7.24 (s, 1 H), 7.15 (m, 1 H), 6 .99 (m, 1 H), 6.94 (m, 1 H), 6.85 (m, 1 H), 6.56 (d, 1 H), 5.93 (s, 2 H), 5 .63 (m, 1 H), 4.77 (m, 1 H), 3.74 (d, 2 H), 0.86 (m, 2 H), 0.65 (m, 2 H).

Stage 2: Synthesis of reference compound I-359

The reference compound I-358 was dissolved in tetrahydrofuran at room temperature and triethylamine (3 equiv.) and ethyl chloroformate (2 equiv.) were added in succession. The content was allowed to stir for 1 hour at room temperature. The mixture was diluted with ethyl acetate, washed with water (3 times), then dried over sodium sulfate, filtered and concentrated in vacuo. The resulting residue was redissolved in tetrahydrofuran, treated with sodium borohydride (6 equiv.), and stirred for 1 h. This mixture was diluted with ethyl acetate, washed with water (3 times), then dried over sodium sulfate, filtered and concentrated in vacuo. The crude material was purified by silica gel chromatography with a gradient of 10-80% ethyl acetate/hexanes providing the desired compound, reference compound I-359 (11 mg, 76% yield) as a white solid.

Reference compound I-360

The title compound was prepared following general procedure B, except that 1-(4-aminophenyl)propanecarboxylic acid (1 equiv.) was the amine reagent, 20 equivalents of triethylamine were used, and the contents were heated to 100° C for 18 h as a solution in dioxane/water (3:1). Ethyl acetate was the solvent used for processing. The crude material was purified by silica gel chromatography with a gradient of 5-95% ethyl acetate/hexanes providing the desired compound, reference compound I-360 (11 mg, 9% yield) as a white solid.

1H-NMR (400 MHz, ace tone^) 58.89 (d, 1 H), 8.81 (br d, 1 H), 8.30 (d, 1 H), 8.03 (d, 2 H ), 7.48 (s, 1 H), 7.39 (d, 2 H), 7.30 (m, 1 H), 7.15 (m, 1 H), 7.10 (m, 1 H ), 7.07 (d, 1 H), 7.05 (m, 1 H), 5.98 (s, 2 H), 1.53 (m, 2 H), 1.18 (m, 2 H ).

Reference compound I-361

The title compound was synthesized in 3 steps:

Step 1: Synthesis of ferf-butyl methyl-(2-oxo-2-(phenylsulfonamido)ethyl)carbamate

To a solution of 1,1-carbonyldiimidazole (1.2 equiv.) in dichloromethane was added 2-((ferf-butoxycarbonyl)(methyl)amino)acetic acid (1 equiv.). The mixture was stirred at 23 °C until gas release ceased. Benzenesulfonamide (3 equiv.) and DBU (1 equiv.) were added to this mixture. The mixture was stirred at 23 °C for 1 h. The mixture was diluted with dichloromethane and washed with water. The organic layer was dried, filtered and evaporated to give a white solid. Purification by silica gel chromatography (gradient from 0 to 80% ethyl acetate in hexanes) afforded ferf-butyl methyl(2-oxo-2-(phenylsulfonamido)ethyl)carbamate (1.3 g) as a white solid. .

1H NMR (500 MHz, CD3OD) 5 ppm 7.91 - 7.95 (m, 2 H), 7.59 - 7.65 (m, 3 H), 3.86 (s, 2 H), 2, 82 - 2.88 (m, 3 H), 1.20 (s, 9 H).

Step 2: Synthesis of 2-(methylamino)-A/-(phenylsulfonyl)acetamide hydrochloride

A mixture of ferf-butyl methyl-(2-oxo-2-(phenylsulfonamido)ethyl)carbamate (1 equiv.) and HCl [4.0 M in 1,4-dioxane] was stirred at 23 °C for 24 h. . The mixture was concentrated to provide 2-(methylamino)-A/-(phenylsulfonyl)acetamide hydrochloride (3.2 g) as a cream solid. 1H NMR (500 MHz, CD3OD) 5 ppm 8.04 - 8.07 (m, 2 H), 7.57 - 7.64 (m, 3 H), 3.88 (s, 2 H), 2, 67 (s, 3 H).

Stage 3: Synthesis of reference compound I-31

The title compound was prepared following general procedure B, except that 2-(methylamino)-N-(phenylsulfonyl)acetamide hydrochloride was the amine reagent, 6 equivalents of triethylamine were used, and the contents were heated to 85° C as a solution in dioxane/water (4:1) for 24 h. The mixture was cooled to 23°C and diluted with ethyl acetate. The organic layer was washed with saturated ammonium chloride solution, dried, filtered and evaporated to give a solid. The solid was purified by silica gel chromatography (gradient from 0 to 100% ethyl acetate in hexanes) providing the desired compound, reference compound I-361 (6.5 mg, 14% yield for step 3) as a light yellow solid.

1H NMR (500 MHz, CD3OD) 5 ppm 8.79 (d, 1 H), 8.12 (d, 1 H), 7.91 - 7.96 (m, 2 H), 7.43 - 7, 49 (m, 1 H), 7.25 - 7.37 (m, 4 H), 7.03 - 7.14 (m, 2 H), 6.95 (d, 1 H), 6.85 - 6.91 (m, 1 H), 5.97 (s, 2 H), 4.30 (s, 2 H), 3.39 (d, 3 H).

Reference compound I-363

The title compound was prepared following general procedure B, except that piperidine-4-sulfonic acid (2 equiv.) was the amine reagent, 2 equivalents of triethylamine were used, and the contents were heated to 70 °C as a solution in dioxane/water (1:1) for 2 h. Ethyl acetate was used as the extraction solvent during preparation. The organic layer was dried, filtered and concentrated in vacuo to provide the desired compound, reference compound I-363 (102 mg, 69% yield) as a white solid.

1 H R M N (500 M H z, D M S O - d a) 5 p p m 9, 12 (d, 1 H), 8, 41 (d, 1 H), 7, 72 (s, 1 H), 7, 28 - 7, 39 (m, 2H), 7, 18 - 7, 27 (m, 1H), 7, 11 (t, 1H), 6, 85 (t, 1H), 5, 95 (s, 2H) , 4 , 65 ( d , 2 H ) , 3 , 21 ( t , 2 H ) , 2 , 61 - 2 , 7 2 ( m , 1 H ) , 2 , 0 2 - 2 , 12 ( m , 2 H ) , 1 , 52 - 1 , 70 ( m , 2 H ) .

Reference compound I-364

The title compound was prepared following general procedure B, except that 4,5,6,7-tetrahydroisoxazolo[5,4-c]pyridin-3-ol (as the HCl salt, 2 equiv.) was the reagent. of amine, 4 equivalents of triethylamine were used, and the content was heated to 70 °C as a solution in dioxane/water (3:1) for 2 h. Ethyl acetate was used as the extraction solvent during preparation. The organic layer was dried, filtered and concentrated in vacuo to give a solid, which was further clarified with diethyl ether and dichloromethane to give the desired compound, reference compound I-364 (46 mg, 72% yield) as a solid. white.

1H NMR (500 MHz, DMSO-d6) 5 ppm 11.47 (s, 1 H), 9.10 (d, 1 H), 8.40 (d, 1 H), 7.62 (s, 1 H) ), 7.30 - 7.37 (m, 1 H), 7.27 (d, 1 H), 7.20 - 7.25 (m, 1 H), 7.11 (t, 1 H), 6.84 (t, 1 H), 5.91 (s, 2 H), 4.90 (s, 2 H), 3.98 (t, 2 H), 2.51 - 2.55 (m, 2H).

Reference compound I-365

The title compound was prepared following general procedure B, except that (1R,3S,4S)-2-azabicyclo[2.2.1]heptane-3-carboxylic acid (as HCl salt, 2 equiv.) was the reagent of amine, 2.5 equivalents of triethylamine were used, and the contents were heated to 60 °C for 24 h, and then to 80 °C for 3 h. Ethyl acetate was used as an extraction solvent during processing. The crude material was purified by silica gel chromatography using a gradient of 0-100% ethyl acetate/hexanes providing a solid, which was further clarified with diethyl ether and dichloromethane providing the desired compound, reference compound I-365 ( 21 mg, yield 16%) as a white solid.

1H NMR (500 MHz, CD3OD) 5 ppm 8.78 (d, 1 H), 8.07 - 8.20 (m, 1 H), 7.23 - 7.35 (m, 2 H), 7, 08 - 7.15 (m, 1 H), 7.04 (t, 1 H), 6.94 (br. s., 1 H), 6.79 - 6.87 (m, 1 H), 5 .92 - 6.01 (m, 2 H), 4.28 - 4.38 (m, 1 H), 2.85 - 2.97 (m, 1 H), 2.23 (d, 1 H) , 1.72 - 1.98 (m, 4 H), 1.48 - 1.71 (m, 2 H).

Reference compound I-366

The title compound was prepared by general procedure B, except that piperidine-4-sulfonamide (2 equiv.) was the amine reagent, 2 equivalents of triethylamine were used, and the contents were heated to 75 °C as a solution. in dioxane/water (3:1) for 2 h. The mixture was diluted with ethyl acetate and washed with 1N HCl solution. The insoluble solids were collected by filtration and dried under vacuum to provide the desired compound, reference compound I-366 (64 mg, 48% yield) as a white solid.

1H NMR (500 MHz, DMSO-d6) 5 ppm 9.10 (d, 1 H), 8.35 (d, 1 H), 7.60 (s, 1 H), 7.31 - 7.37 ( m, 1 H), 7.21 - 7.28 (m, 2 H), 7.11 (t, 1 H), 6.79-6.85 (m, 3 H), 5.91 (s, 2 H), 4.66 (d, 2 H), 3.24 (ddt, 1 H), 3.16 (t, 2 H), 2.13 (d, 2 H), 1.67 (qd, 2H).

Reference compound I-367

The title compound was synthesized in 3 steps:

Stage 1: Synthesis of 2-(1,3-dioxoisoindolin-2-yl)-A/-methylethanesulfonamide.

To a solution of 2-(1,3-dioxoisoindolin-2-yl)-ethanesulfonyl chloride (1 equiv.) in THF was added methylamine (2.0 M solution in THF] (2 equiv.). The mixture was stirred. at 23 °C for 1 h. The mixture was concentrated to give 2-(1,3-dioxoisoindolin-2-yl J-N-methylethylenesulfonamide (0.98 g) as a white solid. 1H NMR (500 M<h z>, CD3OD) 5 ppm 7.79 - 7.92 (m, 4 H), 4.06 - 4.13 (m, 2 H), 3.42 (t, 2 H), 2.74 (s, 3 H).

Step 2: Synthesis of 2-amino-W-methylethylene sulfonamide.

Hydrazine monohydrate (1, 5 equiv.). The mixture was heated at 75 °C for 2 h. The white precipitate formed was removed by filtration. The filtrate was concentrated in vacuo to provide 2-amino-W-methylethanesulfonamide as a white solid.

1H NMR (500 MHz, CD3OD) 8 ppm 3.20 - 3.24 (m, 2 H), 3.08 - 3.14 (m, 2 H), 2.71 (s, 3 H).

Stage 3: Synthesis of reference compound I-367

The title compound was prepared following general procedure B, except that 2-amino-A/-methylethanesulfonamide (2 equiv.) was the amine reagent, 2 equivalents of triethylamine were used, and the contents were heated to 65 °C for 2 d. The mixture was diluted with ethyl acetate and washed with saturated sodium bicarbonate solution. The organic layer was dried, filtered and evaporated to give a solid. The solid was purified by silica gel chromatography (gradient from 0 to 100% ethyl acetate in hexanes) to provide the desired compound, reference compound I-367 (8.6 mg, 14% yield) as a colored solid. cream. 1H NMR (500 MHz, CD3OD) 8 ppm 8.76 (d, 1 H), 8.10 (d, 1 H), 7.49 (s, 1 H), 7.24 - 7.30 (m, 1 H), 7.07 - 7.13 (m, 1 H), 7.03 (t, 1 H), 6.87 - 6.90 (m, 1 H), 6.81 (t, 1 H) ), 5.96 (s, 2 H), 4.01 (t, 2 H), 3.42 (t, 2 H), 2.68 - 2.70 (m, 3 H).

Reference compound I-368

The title compound was prepared following general procedure B, except that W-methyltaurine (as sodium salt, 2 equiv.) was the amine reagent, 2 equivalents of triethylamine were used, and the contents were heated to 65 °C as a solution. in dioxane/water (3:1) for 24 hours. Ethyl acetate was used as a solvent for the preparation. The organic layer was dried, filtered and evaporated to give a solid. The solid was collected by filtration and dried under vacuum to provide the desired compound, reference compound I-368 (31 mg, 33% yield) as a white solid.

1H NMR (500 MHz, DMSO-d6) 8 ppm 9.13 (d, 1 H), 8.40 (d, 1 H), 7.93 (br. s., 1 H), 7.31 - 7 .38 (m, 1 H), 7.21 - 7.27 (m, 1 H), 7.17 (s, 1 H), 7.12 (t, 1 H), 6.88 (t, 1 H), 5.96 (s, 2 H), 3.94 - 4.02 (m, 2 H), 3.38 (d, 3 H), 2.84 - 2.92 (m, 2 H) .

Reference compound I-369

The title compound was prepared following general procedure B, except that c/s-1-amino-1,2,3,4-tetrahydronaphthlene-2-carboxylic acid (2 equiv.) was the amine reagent, 2 equivalents of triethylamine, and the content was heated to 65 °C as a solution in dioxane/water (3:1) for 24 h. Ethyl acetate was used as a solvent for the preparation. The crude material was purified by silica gel chromatography with a gradient of 0-50% ethyl acetate/hexanes to provide the desired compound, reference compound I-369 (7.0 mg, 10% yield) as a white solid. .

1H NMR (500 MHz, DMSO-d6) 8 ppm 9.07 - 9.10 (m, 1 H), 8.22 (d, 1 H), 7.70 (d, 1 H), 7.57 ( s, 1 H), 7.30 - 7.41 (m, 2 H), 7.08 - 7.24 (m, 6 H), 6.89 (t, 1 H), 5.91 (s, 2 H), 2.99 (dt, 1 H), 2.74 - 2.91 (m, 2 H), 2.28 - 2.37 (m, 1 H), 1.94 - 2.01 ( m, 1H).

Reference compound I-370

The title compound was prepared following general procedure B, except that 4,5,6,7-tetrahydro-1H-pyrazolo[3,4-c]pyr¡din-3(2H)-one ( as HCl salt, 3.2 equiv.) was the amine reagent, and the content was heated to 65 °C as a solution in dioxane/water (1:1) for 24 h. The mixture was diluted with ethyl acetate and extracted with 1N HCl solution. The aqueous layer was made basic with saturated sodium bicarbonate solution and extracted with ethyl acetate. The combined organic layers were dried, filtered and evaporated to provide the desired compound, reference compound I-370 (83 mg, 65% yield) as a light brown solid.

1H NMR (500 MHz, CDCh) 8 ppm 8.38 (d, 1 H), 8.08 - 8.21 (m, 1 H), 7.22 (s, 1 H), 7.07 - 7, 13 (m, 1 H), 6.91 (t, 1 H), 6.85 (t, 1 H), 6.73 - 6.78 (m, 1 H), 6.52 (d, 1 H ), 5.85 (s, 2 H), 4.69 (s, 2 H), 3.83 - 3.91 (m, 2 H), 2.52 (t, 2 H).

Reference compound I-371

The title compound was prepared by general procedure B, except that c/s-1-amino-2,3-dihydro-1H-indene-2-carboxylic acid (2 equiv.) was the amine reagent. 2 equivalents of triethylamine were used, and the content was heated to 80 °C as a solution in dioxane/water (3:1) for 24 h. The mixture was diluted with ethyl acetate and washed with 1N HCl solution. The organic layer was dried, filtered and concentrated in vacuo. The resulting solid was clarified with trace amounts of methanol to provide the desired compound, reference compound I-371 (11 mg, 16% yield) as a white solid.

1H NMR (500 MHz, DMSO-d6) 8 ppm 9.10 (d, 1 H) 8.25 (d, 1 H) 7.63 (s, 1 H) 7.29 - 7.39 (m, 3 H) 7.20 - 7.28 (m, 3 H) 7.09 - 7.19 (m, 2 H) 6.88 (t, 1 H) 6.22 - 6.29 (m, 1 H) 5.87 - 5.97 (m, 2 H) 3.67 (q, 1 H) 3.52 (dd, 1 H) 3.06 (dd, 1 H).

Reference compound I-372

The composition of the title was synthesized in 3 stages:

Stage 1: Synthesis of ferf-butyl methyl(2-oxo 2-(phenylsulfonamido)ethyl)carbamate.

To a suspension of 2-(((ferf-butoxycarbonyl)amino)methyl)thiazole-4-carboxylic acid (1 equiv.) and methyl iodide (10 equiv.) in THF at 0 °C s was added sodium hydride [dispersion at 60% by weight in mineral oil] (10 equiv.). The mixture was stirred at 23 °C for 24 h. The mixture was diluted in ethyl acetate and washed with 1N HCl solution. The organic layer was dried, filtered and evaporated to give an oil, which was further purified by silica gel chromatography (0-50% ethyl acetate/hexanes gradient) to provide 2-(((ferfbutoxycarbonyl)(methyl)amino acid. )methyl)thiazole-4-carboxylic acid (217 mg, yield 82%) as a red solid.

1H NMR (500 MHz, CDCla) 8 ppm 8.26-8.34 (m, 1 H), 4.76 (d, 2 H), 3.00 (d, 3 H), 1.51 (br. s, 9H).

Stage 2: Synthesis of ethyl 2-((methylamino)methyl)thiazole-4-carboxylate hydrochloride.

A mixture of 2-(((ferf-butoxycarbonyl)(methyl)amino)methyl)thiazole-4-carboxylic acid (1 equiv.) and a solution of HCl [1.3 M in ethanol] (10 equiv.) was stirred at 23 °C for 24 h. The mixture was concentrated to provide ethyl 2-((methylamino)methyl)thiazole-4-carboxylate (as HCl salt, 222 mg) as a yellow solid.

1H NMR (500 MHz, CD3OD) 8 ppm 8.46 (s, 1 H), 4.63 (s, 2 H), 4.27-4.37 (m, 2 H), 2.80 (s, 3 H), 1.28- 1.35 (m, 3 H). Stage 3: Synthesis of reference compound I-372

The title compound was prepared following general procedure B, except that ethyl 2-((methylamino)methyl)thiazole-4-carboxylate (as the HCl salt, 2 equiv.) was the amine reagent, 6 were used equivalents of triethylamine, and the content was heated to 80 °C as a solution in dioxane/water (4:1) for 24 h. The mixture was diluted in ethyl acetate and washed with saturated sodium bicarbonate solution. The organic layer was dried, filtered and evaporated to give a solid, which was purified by silica gel chromatography (gradient from 0 to 100% ethyl acetate in hexanes) to provide the desired compound, reference compound I-372 (78 mg, yield 77%) as a light yellow gum.

1H NMR (500 MHz, CDCla) 8 ppm 8.45 (d, 1 H), 8.26 (d, 1 H), 8.12 (s, 1 H), 7.28 (s, 1 H), 7.17 - 7.23 (m, 1 H), 6.94 -7.06 (m, 2 H), 6.86 - 6.92 (m, 1 H), 6.58 (d, 1 H ), 5.97 (s, 2 H), 5.22 (s, 2 H), 4.44 (q, 2 H), 3.42 (d, 3 H), 1.42 (t, 3 H ). Reference compound I-373

A mixture of reference compound I-372 (1 equiv.) and lithium hydroxide (10 equiv.) in a THF/water/methanol mixture (1:1:1) was stirred at 23 °C for 24 h. The mixture was concentrated in vacuo. The resulting solution was acidified to pH 1. The precipitate formed was collected by filtration and dried under vacuum to provide the desired compound, reference compound I-373 (38 mg, 67% yield) as a white solid.1

1H NMR (400 MHz, DMSO-cfe) 8 ppm 13.05 (s, 1 H), 9.09 (d, 1 H), 8.36 (t, 2 H), 7.57 (s, 1 H) ), 7.30 - 7.37 (m, 1 H), 7.18 - 7.26 (m, 2 H), 7.11 (t, 1 H), 6.89 (t, 1 H), 5.89 (s, 2 H), 5.17 (s, 2 H), 3.32-3.39 (m, 3 H).

Reference compound I-374

A suspension of reference compound I-368 (1 equiv.) in thionyl chloride (50 equiv.) was heated at reflux for 24 h. The mixture was cooled and concentrated in vacuo to give the sulfonyl chloride intermediate 2-((5-fluoro-2-(1-(2-fluorobenzyl)-5-(isoxazol-3-yl)-1H-chloride). pyrazol-3-yl)pyrimidin-4-yl)(methyl)amino)-ethanesulfonyl (169 mg) as a yellow solid. This material was then treated with ammonia [0.05 M solution in 1,4-dioxane] (5 equiv.) and the mixture was allowed to stir at 23 °C for 24 h. The reaction was diluted in ethyl acetate and washed with saturated sodium bicarbonate solution. The organic layer was dried, filtered and evaporated to give an oil. The oil was purified by silica gel chromatography (gradient from 0 to 30% methanol in dichloromethane) and recrystallized from a methanol:dichloromethane mixture to provide the desired compound, reference compound I-374 (13 mg, yield 8%) as a light yellow solid.

1 H R M N (400 M H z, D M S O -d 6) 8 ppm 9 .10 (d, 1 H) 8 .28 (d, 1 H) 7 .58 (s, 1 H) 7 .33 (d, 1 H) 7 .19 - 7 .25 (m, 1 H) 7 .14 (d, 1 H) 7 .08 - 7 .13 (m, 2 H) 5 .89 (s, 2 H) 4 .00 - 4 , 06 (m, 2 H) 3 .36 - 3.41 (m, 2 H) 3 .28 (d, 3 H).

I-375 REFERENCE COMPOSITION

The title composition was prepared in 3 stages:

Stage 1: Synthesis of 2-(((terf-butoxy¡carbon¡l)(methyl)amino)methyl)oxazole-4-carboxylic acid

To a cold suspension of 2-(((terf-butoxy¡carbon¡l)amino)methyl)oxazole-4-carboxylic acid (1 equiv.) and iodomethane (10 equiv. v.) in THF at 0 °C, sodium hydride [dispersion at 60% by weight in mineral oil] (10 equiv.) was added. The mixture was allowed to warm to 23°C and stirred for 24 h. The mixture was diluted in ethyl acetate and washed with 1N HCl solution. The organic layer was dried, filtered and evaporated giving an oil. The oil was purified by column chromatography (gradient from 0 to 100% ethyl acetate in hexanes) providing 2-(((tert-butoxycarbonyl)( methyl)amino)methyl)oxazole-4-carboxylic acid (215 mg, yield 81%) as a yellow oil.

1H NMR (400 MHz, CDCla) 8 ppm 8.27 (s, 1 H), 4 .52-4.69 (m, 2 H), 2.93 (d, 3 H), 1.37- 1, 53 (m, 9 H).

Stage 2: Syntheses of 2-((methylamino)methyl)oxazole-4-carboxylic acid, TFA salt.

A mixture of 2-(((terf-butoxycarbon¡l)(methyl)amino)methyl)oxazole-4-carboxylic acid (1 eq) and TFA (10 eq v.) in DCM was stirred at 23 °C for 1 h. The mixture was concentrated to provide 2-((methylamino)methyl)oxazole-4-carboxylic acid, TFA salt (237 mg) as a clear oil. 1H R<m>N (500 MHz, CD3OD) 8 ppm 8.62 - 8.64 (m, 1 H), 4.49 (s, 2 H), 2.86 (s, 3 H).

Stage 3: Syntheses of the reference compound I-375

The title compound was prepared following general procedure B, except that 2-((methlano)methyl)oxazole-4-carboxylic acid (as salt of TFA, 4 equiv.) was the amine reagent, 8 equivalents of triethylamine were used and the content was heated to 80 °C as a solution in dioxane/water (4:1) for 24 hours. Ethyl acetate was used as a solvent during preparation. The crude oil obtained after preparation was treated with diethyl ether and the resulting precipitated solid was collected by filtration and dried under vacuum proportional to ionizing the desired compound, the reference compound I-375 (yield 68%) as a white solid.

1H NMR (500 MHz, DMSO-d6) 8 ppm 9.10 (d, 1 H), 8.73 (s, 1 H), 8.36 (d, 1 H), 7.50 (s, 1 H) ), 7.28 - 7.36 (m, 1 H), 7.16 - 7.26 (m, 2 H), 7.10 (t, 1 H), 6.85 (t, 1 H), 5.88 (s, 2 H), 5.07 (s, 2 H), 3.35 - 3.40 (m, 3 H).

Reference compound I-376

The title compound was synthesized in 2 steps:

Stage 1: Syntheses of 1-methyl-2-((methylamino)methyl)-1H-midazole-4-carboxylic acid hydrochloride.

To a suspension of 2-(((tert-butoxycarbonyl)amino)methyl)-1H-midazole-4-carboxylic acid (1 equiv.) and methyl iodide (10 equiv.) in THF at 0 °C sodium hydride was added [dispersion at 60% by weight in mineral oil] ( 10 equiv.). The mixture was stirred at 23°C for 24 h. The mixture was treated with HCl solution [4.0 M solution in 1,4-dioxane]. The mixture was concentrated in vacuo and the resulting solid was suspended in diethyl ether. The precipitate was collected by filtration and dried in vacuo giving the acid 1-methyl-2-((methylamino)metal)-1H-m dazole-4-carboxylic acid (80 mg, yield 38%) as a yellow solid.

1H NMR (500 MHz, DMSO-d6) 8 ppm 9.34 (br. s., 2 H), 7.91 (s, 1 H), 4.29 (t, 2 H), 3.71 (s , 3 H), 2.64 (t, 3 H).

Step 2: Synthesis of the reference compound I-376

The title compound was prepared following general procedure B, except that 1-methyl-2-((methylamino)methyl)-1H-imidazole-4-carboxylic acid (as the HCl salt, 3 equiv.) was the reagent. of amine, 4 equivalents of triethylamine were used, and the content was heated to 80 °C as a solution in dioxane/water (4:1) for 24 h. The mixture was cooled to 23 °C, diluted in ethyl acetate and washed with 1N HCl solution. The precipitate formed was collected by filtration and dried under vacuum to provide the desired compound, reference compound I-376 (26 mg, 37% yield) as a white solid.

1H NMR (500 MHz, DMSO-cfe) 8 ppm 9.10 (d, 1 H), 8.39 (d, 1 H), 8.11 (br. s., 1 H), 7.56 (s , 1 H), 7.28 - 7.39 (m, 1 H), 7.18 - 7.26 (m, 2 H), 7.07 - 7.14 (m, 1 H), 6.82 (t, 1 H), 5.91 (s, 2 H), 5.08 (s, 2 H), 3.85 (s, 3 H), 3.41 (d, 3 H).

Reference compound I-377

The title compound was prepared following general procedure B, except that 3-(2-aminoethyl)benzoic acid was the amine reagent, 4 equivalents of triethylamine were used, and the contents were heated to 90°C as a solution in dioxane/water (3:1) for 24 h. Ethyl acetate was used as a solvent during preparation. The crude material was purified by silica gel chromatography with a gradient of 0-80% ethyl acetate/hexanes providing the desired compound, reference compound I-377 (25 mg, 31% yield) as a white solid.

1H NMR (500 MHz, CDCh) 8 ppm 8.44 (d, 1 H), 8.17 (d, 1 H), 8.04 (s, 1 H), 7.98 (d, 1 H), 7.50 (d, 1 H), 7.40 - 7.45 (m, 1 H), 7.37 (s, 1 H), 7.14 - 7.21 (m, 1 H), 7, 00 (t, 1 H), 6.92 - 6.97 (m, 1 H), 6.85 - 6.92 (m, 1 H), 6.64 (d, 1 H), 5.99 ( s, 2 H), 5.24 - 5.31 (m, 1 H), 3.90 - 3.97 (m, 2 H), 3.08 (t, 2 H).

Reference compound I-378

The title compound was prepared following general procedure B, except that 5-hydroxy-1,2,3,6-tetrahydropyridine-4-carboxylate (as TFA salt, 1 equiv.) was the amine reagent, 3 equivalents of triethylamine and the content was heated to 70 °C as a solution in dioxane/water (3:1) for 24 h. Ethyl acetate was used as a solvent during preparation. The crude material was purified by silica gel chromatography with a gradient of 0-30% ethyl acetate/hexanes providing the desired compound, reference compound I-378 (68 mg, 15% yield) as a white solid.

1H NMR (500 MHz, CDCla) 8 ppm 12.17 (s, 1 H), 8.46 (d, 1 H), 8.23 (d, 1 H), 7.31 (s, 1 H), 7.16 - 7.22 (m, 1 H), 7.02 (t, 1 H), 6.96 (t, 1 H), 6.84 (t, 1 H), 6.62 (d, 1 H), 5.98 (s, 2 H), 4.47 - 4.50 (m, 2 H), 4.26 (q, 2 H), 3.94 (t, 2 H), 2, 51 (t, 2 H), 1.33 (t, 3 H).

Reference compound I-379

The title compound was prepared following general procedure B, except that (1SR,2SR,3RS,4RS)-3-aminobicyclo[2.2.1]heptane-2-carboxylic acid (as the HCl salt, racemic, 3 equiv. ) was the amine reagent, 6 equivalents of triethylamine were used, and the content was heated to 70 °C as a solution in dioxane/water (3:1) for 24 h. The reaction mixture was cooled, diluted with ethyl acetate and washed with water. The organic layer was dried, filtered and concentrated in vacuo. The crude material was purified by silica gel chromatography with a gradient of 0-80% ethyl acetate/hexanes providing the desired compound, reference compound I-379 (47 mg, 36% yield) as a white solid.

1H NMR (500 MHz, CD3OD) 8 ppm 8.76 - 8.79 (m, 1 H), 8.07 (d, 1 H), 7.42 (s, 1 H), 7.25 - 7, 32 (m, 1 H), 7.11 (ddd, 1 H), 7.05 (td, 1 H), 6.92 (d, 1 H), 6.83 (td, 1 H), 5, 97 (s, 2 H), 4.58-4.66 (m, 1 H), 3.13 (ddd, 1 H), 2.83 (br. s., 1 H), 2.67 (br s., 1 H), 1.74 (d, 1 H), 1.65 - 1.71 (m, 1 H), 1.58 - 1.64 (m, 1 H), 1.49 - 1.57 (m, 2 H), 1.41 - 1.49 (m, 1 H).

Reference compound I-380

The title compound was prepared following general procedure B, except that (1S,3R)-3-aminocyclohexanecarboxylic acid was the amine reagent (98% ee, 4 equiv.), 4 equivalents of triethylamine were used, and the content was heated to 80 °C as a solution in dioxane/water (3:1) for 24 h. Ethyl acetate was used as a solvent during preparation. The crude material was purified by silica gel chromatography with a gradient of 0-10% methanol/dichloromethane providing the desired compound, reference compound I-380 (70 mg, 55% yield) as a yellow solid.

1H NMR (500 MHz, CD3OD) 8 ppm 8.81 (d, 1 H), 8.18 (d, 1 H), 7.59 (s, 1 H), 7.29 (q, 1 H), 7.07 - 7.14 (m, 1 H), 7.05 (t, 1 H), 6.98 (d, 1 H), 6.92 (t, 1 H), 6.00 (s, 2 H), 4.46 (t, 1 H), 2.59 (t, 1 H), 2.31 (d, 1 H), 2.04 (d, 2 H), 1.95 (d, 1 H), 1.51 -1.65 (m, 2 H), 1.35-1.51 (m, 2 H).

Reference compound I-381

The composition of the title was synthesized in 3 stages:

Step 1: Synthesis of 2-(1,3 dioxoisoindolin-2-yl )-N-(2,2,2-trifluoroethyl)ethanesulfonamide

A mixture of 2-[2-(chlorosulfonyl)ethyl]benzo[c]azolin-1,3-dione (1 equiv.), 2,2,2-trifluoroethylamine hydrochloride (3 equiv.), and triethylamine (6 equiv. ) in dichloromethane was stirred at 23 °C for 24 h. The mixture was diluted in dichloromethane and washed with 1N HCl solution. The organic layer was dried, filtered and evaporated to give a white solid. The solid was purified by silica gel chromatography (0 to 100% ethyl acetate in hexanes) giving 2-(1,3-dioxoisoindolin-2-yl)-N-(2,2,2-trifluoroethyl)ethanesulfonamide ( 370 mg, yield 30%) as a white solid.

1H NMR (500 MHz, DMSO-da) 8 ppm 7.84 - 7.92 (m, 4 H), 4.05 - 4.09 (m, 2 H), 3.97 - 4.01 (m, 2H), 3.32 (s, 2H).

Step 2: Synthesis of 2-amino-N-(2,2,2-trifluoroethyl)ethanesulfonamide

A mixture of 2-(1,3-dioxoisoindolin-2-yl)-N-(2,2,2-trifluoroethyl)ethanesulfonamide (1 equiv.) and hydrazine monohydrate (1 equiv.) in ethanol was stirred at 80° C for 24 hours. The mixture was cooled to 23 °C and concentrated in vacuo. The resulting residue was treated with a minimum amount of methanol. The precipitate was collected by filtration and dried under vacuum to provide a white solid containing 2-amino-N-(2,2,2-trifluoroethyl)ethanesulfonamide (136 mg). The material was used in the following reaction without further purification.

Stage 3: Synthesis of reference compound I-381

The title compound was prepared following general procedure B, except that 2-amino-N-(2,2,2-trifluoroethyl)ethanesulfonamide (1 equiv.) was the amine reagent, and the contents were heated to 80° C as a solution in dioxane/water (3:1) for 24 h. The mixture was cooled, diluted in ethyl acetate and washed with water. The organic layer was dried, filtered and evaporated to give an oil. The oil was purified by silica gel chromatography (gradient from 0 to 70% ethyl acetate in hexanes) providing the desired compound, reference compound I-361 (28 mg, 14% yield for step 2 and 3) as a white solid. 1H NMR (500 MHz, DMSO-cfe) 8 ppm 9.10 (d, 1 H), 8.25 (d, 2 H), 7.55 (s, 1 H), 7.30 - 7.37 ( m, 1 H), 7.19 - 7.26 (m, 1 H), 7.15 (d, 1 H), 7.10 (t, 1 H), 6.82 (t, 1 H), 5.89 (s, 2 H), 3.87 - 3.82 (m, 4 H), 3.45 (t, 2 H).

Reference compound I-382

The title compound was synthesized in 2 steps:

Stage 1: Synthesis of 2-hydroxy-N-2-dimethyl-3-(methylamino)propanamide

In a sealed vial, a mixture of methyl 2-methylglycidate (1 equiv.) and methylamine [33 wt % in THF] (10 equiv.) was heated at 80 °C for 24 h. The mixture was concentrated in vacuo to give 2-hydroxy-N-2-dimethyl-3-(methylamino)propanamide (1.7 g, 100% yield) as a clear oil. 1H NMR (500 MHz, CDCh) 8 ppm 3.67 - 3.76 (m, 1 H), 3.22 - 3.29 (m, 1 H), 2.79 - 2.82 (m, 3 H) ), 2.40 - 2.43 (m, 3 H), 1.31 -1.33 (m, 3 H).

Stage 2: Synthesis of reference compound I-382

The title compound was prepared following general procedure B, except that 2-hydroxy-N-2-dimethyl-3-(methylamino)propanamide (4 equiv.) was the amine reagent, 4 equivalents of triethylamine were used, and the Content was heated to 80 °C as a solution in dioxane/water (3:1) for 4 h. Ethyl acetate was used as a solvent during preparation. The resulting solid was treated with a minimum amount of methanol and diethyl ether, collected by filtration and dried under vacuum to provide the desired compound, reference compound I-382 (67 mg, 52% yield) as a white solid.

1H NMR (500 MHz, DMSO-da) 5 ppm 9.09 - 9.12 (m, 1 H), 8.23 (d, 1 H), 7.87 (q, 1 H), 7.54 ( s, 1 H), 7.30 - 7.37 (m, 1 H), 7.18-7.26 (m, 1 H), 7.11 (td, 1 H), 6.88 (t, 1 H), 5.86-5.92 (m, 2 H), 4.12 (d, 1 H), 3.68 (d, 1 H), 3.22 (s, 3 H), 2, 55 (d, 3 H), 1.27 (s, 3 H).

Reference compound I-383

The title compound was synthesized in 5 steps:

Step 1: Synthesis of benzyl (3-hydroxy-2,2-dimethylpropyl)carbamate

To a cold mixture of 3-amino-2,2-dimethyl-1-propanol (1 equiv.) and triethylamine (1 equiv.) in dichloromethane was added benzyl chloroformate (1 equiv.). The mixture was stirred at 23 °C for 30 min, then diluted in dichloromethane and washed with 1N HCl solution. The organic layer was dried, filtered and evaporated to give benzyl (3-hydroxy-2,2-dimethylpropyl)carbamate (1.9 g, 86% yield) as a white solid.

1H NMR (500 MHz, CDCla) 5 ppm 7.34 - 7.45 (m, 5 H), 5.14 (s, 2 H), 3.25 (d, 2 H), 3.08 (d, 2H), 0.89 (s, 6H).

Step 2: Synthesis of 3-(((benzyloxy)carbonyl)amino)-2,2-dimethylpropanoic acid

To a suspension of benzyl (3-hydroxy-2,2-dimethylpropyl)carbamate (1 equiv.) in CCl4, water and acetonitrile (1:1:1 mixture) sodium periodate (3 equiv.) and sodium chloride were added. ruthenium (III) (0.05 equiv.). The mixture was stirred at 23 °C for 24 h. Additional amounts of sodium periodate (3 equiv.) and ruthenium(III) chloride (0.05 equiv.) were added to this mixture, and the contents were stirred at 23 °C for 3 days. The mixture was diluted with water and extracted with ethyl acetate. The organic layer was extracted with saturated sodium carbonate solution, then the aqueous layer was acidified to pH 1 and extracted with ethyl acetate. The organic layer was dried, filtered and evaporated to give an oil. The oil was purified by silica gel chromatography (gradient from 0 to 100% ethyl acetate in hexanes) to give 3-(((benzyloxy)carbonyl)amino)-2,2-dimethylpropanoic acid (943 mg, 45% yield). like a red oil.

1H NMR (500 MHz, CDCla) 5 ppm 7.35- 7.42 (m, 5 H), 5.12 (s, 2 H), 3.35 (d, 2 H), 1.23- 1, 28 (m, 6 H).

Step 3: Synthesis of 3-(((benzyloxy)carbonyl)(methyl)amino)-2,2 dimethylpropanoic acid

To a cold solution of 3-(((benzyloxy)carbonyl)amino)-2,2-dimethylpropanoic acid (1 equiv.) in THF was added sodium hydride [60 wt.% dispersion in mineral oil] (10 equiv. ). The mixture was stirred at 0 °C for 30 min. Methyl iodide (10 equiv.) was added to this mixture. The mixture was allowed to warm to 23 °C and stirred for 24 h. The mixture was poured onto ice and acidified to pH 1. The mixture was extracted with diethyl ether, and the organic layer was dried, filtered and evaporated to give an oil. The oil was purified by silica gel chromatography (gradient from 0 to 80% ethyl acetate in hexanes) to give 3-(((benzyloxy)carbonyl)(methyl)amino)-2,2-dimethylpropanoic acid (686 mg, yield 69%) as a transparent oil.

1H NMR (500 MHz, CDCh) 5 ppm 7.36 - 7.39 (m, 4 H), 7.32 - 7.35 (m, 1 H), 5.13 (s, 2 H), 3, 53 - 3.60 (m, 2 H), 2.96 (br. s., 3 H), 1.18-1.26 (m, 6 H).

Step 4: Synthesis of 2,2-dimethyl-3-(methylamino)propanoic acid hydrochloride

A solution containing 3-(((benzyloxy)carbonyl)(methyl)amino)amino)-2,2-dimethylpropanoic acid (1 equiv.) in methanol was hydrogenated using the H-cube (0.7 ml/min, catalyst : 10% Pd/C, 70 °C). The resulting mixture was treated with a HCl solution [1.25 M in ethanol] and concentrated in vacuo to give 2,2-dimethyl-3-(methylamino)propanoic acid hydrochloride (569 mg, 100% yield) as an oil. transparent.1

1H NMR (500 MHz, CD3OD) 5 ppm 3.09 (s, 2 H), 2.69 - 2.74 (m, 3 H), 1.25 -1.31 (m, 6 H).

Step 5: Synthesis of the reference compound I-383

The title compound was prepared following general procedure B, except that 2,2-dimethyl-3-(methylamino)propanoic acid (as HCl salt, 4 equiv.) was the amine reagent, 8 equivalents of triethylamine were used and the content was heated to 80 °C as a solution in dioxane/water (3:1) for 24 hours. Ethyl acetate was used as a solvent during preparation. The crude material was purified by silica gel chromatography (gradient from 0 to 60% ethyl acetate in hexanes) to provide the desired compound, reference compound I-383 (38 mg, 30% yield) as a white solid.

1H NMR (500 MHz, DMSO-da) 8 ppm 12.51 (br. s., 1 H), 9.10 (d, 1 H), 8.25 (d, 1 H), 7.49 (s , 1 H), 7.30 - 7.37 (m, 1 H), 7 .18-7.25 (m, 2 H), 7.11 (td, 1 H), 6.92 (t, 1 H), 5.88 (s, 2 H), 3.96 (s, 2 H), 3.33 (s, 3 H), 1.13 (s, 6 H).

Reference compound I-384

The title compound was prepared following general procedure B, except that (1RS,2SR,3RS,4SR)-3-amino-7-oxabicyclo[2.2.1]heptane-2-carboxylic acid (i.e., racemic, 4 equiv.) was the amine reagent, 4 equivalents of triethylamine were used, and the content was heated to 80 °C as a solution in dioxane/water (3:1) for 24 h. Ethyl acetate was used as a solvent during preparation. The resulting solid was clarified with methanol and diethyl ether, collected by filtration and dried under vacuum to provide the desired compound, reference compound I-384 (43 mg, 33% yield) as a white solid.

1H NMR (500 MHz, DMSO-d6) 8 ppm 12.40 (br. s., 1 H), 9.10 (d, 1 H), 8.23 (d, 1 H), 7.54 (s , 1 H), 7.30 - 7.36 (m, 1 H), 7.20 - 7.25 (m, 2 H), 7.09 - 7.13 (m, 1 H), 6.94 (d, 1 H), 6.83 - 6.87 (m, 1 H), 5.89 (s, 2 H), 4.86 (t, 1 H), 4.72 (d, 1 H) , 4.47 (d, 1 H), 3.10 (d, 1 H), 1.53 - 1.68 (m, 4 H).

Reference compound I-436

The title compound was prepared following general procedure B, except that 2-aminoethanesulfonamide was the amine reagent, 3 equivalents of triethylamine were used, and the contents were heated to 65 °C for 24 h as a solution in dioxane. The mixture was diluted in ethyl acetate and washed with saturated ammonium chloride solution. The organic layer was dried, filtered and evaporated to give a solid. The solid was purified by silica gel chromatography (0-100% ethyl acetate in hexanes gradient) to provide the desired compound, reference compound I-436 (26 mg, 42% yield) as a white solid.

1H NMR (500 MHz, CD3OD) 8 ppm 8.77 (d, 1 H), 8.12 (d, 1 H), 7.50 (s, 1 H), 7.25 - 7.32 (m, 1 H), 7.03 - 7.14 (m, 2 H), 6.90 (d, 1 H), 6.82 - 6.88 (m, 1 H), 5.98 (s, 2 H) ), 4.07 - 4.15 (m, 5 H), 3.47 (t, 2 H).

Reference compound I-385

CDI (10 equiv.) was added to a solution of acetic acid (10 equiv.) in DMF. The mixture was stirred at 45 °C for 30 min. The reference compound I-436 and DBU (10 equiv.) were added to this mixture. The mixture was stirred at 23 °C for 3 d. The mixture was diluted in ethyl acetate and washed with 1N HCl solution. The organic layer was dried, filtered and evaporated to give an oil. The oil was purified by silica gel chromatography (gradient from 0 to 10% methanol in dichloromethane) to provide the desired compound, reference compound I-385 (8.2 mg, 25% yield) as a white solid.

1H NMR (500 MHz, DMSO-d6) 8 ppm 9.10 (d, 1 H), 8.24 (d, 1 H), 7.55 (s, 1 H), 7.30 - 7.37 ( m, 1 H), 7.17 - 7.26 (m, 2 H), 7.11 (t, 1 H), 6.86 (t, 1 H), 5.90 (s, 2 H), 3.75-3.87 (m, 4 H), 1.99 (s, 3 H).

Reference compound I-387

The title compound was synthesized in 3 steps:

Step 1: Synthesis of (1-(5-fluoro-2-(1-(2-fluorobenzyl)-5-(isoxazol-3-yl)-1H-pyrazol-3-yl)pyrimidin-4-yl)acetidin- tert-butyl 3-yl)carbamate (Reference Compound I-21)

The intermediate title compound was prepared following general procedure B, except that 3-(tert-butoxycarbonylamino)acetidine was the amine reagent, 3 equivalents of triethylamine were used, and the contents were heated to 80 °C as a solution in dioxane. /water (3:1) for 24 hours. Ethyl acetate was used as a solvent during preparation. The organic layer was dried, filtered and concentrated in vacuo to provide the desired intermediate, 1-(5-fluoro-2-(1-(2-fluorobenzyl)-5-(isoxazol-3-yl)-1H-pyrazole. Ferf-butyl -3-yl)pyrimidin-4-yl)azetidin-3-yl)carbamate (480 mg, 100% yield) as a white solid.

1H NMR (500 MHz, DMSO-d6) 8 ppm 9.09 (d, 1 H), 8.28 (d, 1 H), 7.65 (d, 1 H), 7.53 (s, 1 H) ), 7.29 - 7.35 (m, 1 H), 7.20 - 7.25 (m, 2 H), 7.08 - 7.12 (m, 1 H), 6.81 (t, 1 H), 5.91 (s, 2 H), 4.49 (br. s., 3 H), 4.12 (br. s., 2 H), 1.40 (s, 9 H).

Step 2: Synthesis of 1-(5-fluoro-2-(1-(2-fluorobenzyl)-5-(isoxazol-3-yl)-1H-pyrazol-3-yl)pyrimidin-4-yl)azetidin-3 -amine (Reference compound I-40)

A mixture of (1 -(5 -flu o ro -2 -(1 -(2 -flu o ro benzyl)-5 -( is o x a z o l -3 -il) -1 H -p ir a z o l -3 - il)p ir im id in -4 - il)a z e tid in -3 - il)c a rb a m a to of te rt-butyl (1 eq u iv .) and T F A (10 eq u iv .) in di chlo ro m e tan o was stirred at 23 °C for 24 h. The mixture was concentrated in vacuo. The resulting oil was treated with saturated sodium bicarbonate solution and extracted with ethyl acetate. The precipitate formed was collected and dried under vacuum. The organic layer was also dried, filtered and evaporated to give 1-(5-fluoro-2-(1-(2-fluorobenzyl)-5-(isoxazol-3-yl)-1H-pyrazol-3-yl)pyrimidin. -4-yl)azetidin-3-amine (116 mg (combined), 36% yield) as a white solid.

1H NMR (500 MHz, CD3OD) 8 ppm 8.81 - 8.83 (m, 1 H) 8.32 (d, 1 H) 7.52 - 7.55 (m, 1 H) 7.27 - 7 .34 (m, 1 H) 7.02 -7.14 (m, 2 H) 6.89 - 6.95 (m, 2 H) 6.00 (s, 2 H) 4.84 - 4.88 (m, 2 H) 4.55 (d, 2 H) 4.35 - 4.42 (m, 1 H).

Stage 3: Synthesis of reference compound I-387

To a white suspension containing 1-(5-fluoro-2-(1-(2-fluorobenzyl)-5-(isoxazol-3-yl)-1H-pyrazol-3-yl)pyrimidin-4-yl)azetidin- 3-amine (1 equiv.) and pyridine (10 equiv.) in THF trifluoromethanesulfonic anhydride (2 equiv.) was added. The mixture was stirred at 23 °C for 30 min. The mixture was diluted with ethyl acetate and washed with 1N HCl solution. The organic layer was dried, filtered and evaporated to give an oil. The oil was purified by silica gel chromatography (gradient from 0 to 80% ethyl acetate in hexanes) to provide the desired compound, reference compound I-387 (61 mg, 48% yield) as a white solid.

1H NMR (500 MHz, DMSO-cfe) 8 ppm 10.10 (d, 1 H), 9.08 (d, 1 H), 8.30 (d, 1 H), 7.52 (s, 1 H) ), 7.30 - 7.36 (m, 1 H), 7.16 - 7.27 (m, 2 H), 7.10 (t, 1 H), 6.81 (t, 1 H), 5.90 (s, 2 H), 4.71 - 4.83 (m, 1 H), 4.52 - 4.63 (m, 2 H), 4.27 (dd, 2 H).

Reference compound I-388

To a cold suspension of 1-(5-fluoro-2-(1-(2-fluorobenzyl)-5-(isoxazol-3-yl)-1H-pyrazol-3-yl)pyrimidin-4-yl)azetidin-3 -amine (generated in step 2 of the procedure for the reference compound I-387 1 equiv.) and pyridine (4 equiv.) in dichloromethane at -78 ° C, trifluoromethanesulfonic anhydride (2 equiv.) was added. The mixture was stirred at -78 °C for 2 h. It was then removed from the dry ice acetone bath and heated to 23°C. Stirring was continued at 23 °C for 1 h. The mixture was quenched with methanol, and concentrated in vacuo. The residue was dissolved in ethyl acetate and washed with 1N HCl solution. The organic layer was dried, filtered and evaporated to give an oil. The oil was purified by silica gel chromatography (gradient from 0 to 100% ethyl acetate in hexanes) and the isolated material was recrystallized from a mixture of diethyl ether and methanol providing the desired compound, reference compound I- 388 (90 mg, 19% yield) as a light yellow solid.

1H NMR (500 MHz, DMSO-d6) 8 ppm 9.10 (d, 1 H), 8.35 (br. s., 1 H), 7.57 (br. s., 1 H), 7, 31 - 7.37 (m, 1 H), 7.19 - 7.28 (m, 2 H), 7.11 (t, 1 H), 6.82 (t, 1 H), 5.92 ( s, 2 H), 4.67 (br. s., 2 H), 4.61 (br. s., 1 H), 4.22 (br. s., 2 H).

Reference compound I-389

The title compound was synthesized in 2 steps:

Stage 1: Synthesis of 3-amino-2-hydroxy-2-methylpropanamide

A mixture of ammonia [7 M in methanol] (20 equiv.) and methyl 2-methylglycidate (1 equiv.) was stirred in a sealed vial at 80 °C for 24 h. The mixture was concentrated in vacuo giving 3-amino-2-hydroxy-2-methylpropanamide (731 mg, 100% yield) as a clear oil which transformed into a white solid when left at 23 °C.

1H NMR (500 MHz, CD3OD) 8 ppm 2.90 - 2.96 (m, 1 H) 2.60-2.66 (m, 1 H) 1.32- 1.35 (m, 3 H).

Stage 2: Synthesis of reference compound I-389

The title compound was prepared following general procedure B, except that 3-amino-2-hydroxy-2-methylpropanamide (4 equiv.) was the amine reagent, 4 equivalents of triethylamine were used, and the contents were heated to 80 °C as a solution in dioxane/water (3:1) for 24 h. The mixture was diluted in ethyl acetate and washed with water. The organic layer was dried, filtered and evaporated to give a solid. The solid was purified by silica gel chromatography (gradient from 0 to 5% methanol in dichloromethane) to provide the desired compound, reference compound I-389 (78 mg, 25% yield) as a white solid.

1H NMR (500 MHz, DMSO-cfe) 8 ppm 8.23 (d, 1 H), 7.50 (s, 1 H), 7.30 - 7.35 (m, 2 H), 7.27 ( t, 1 H), 7.25 - 7.29 (m, 1 H), 7.20 - 7.24 (m, 3 H), 7.10 (td, 1 H), 6.86 (t, 1 H), 5.94 (s, 1 H), 5.89 (s, 2 H), 3.75 (dd, 1 H), 3.59 (dd, 1 H), 1.28 (s, 3H).

Reference compound I-390

The title compound was prepared by following the general procedure B, except that the W -(2-aminoethyl)carb am a of fe d -butyl (4 eq u iv .) was the amine reagent, 4 eq uiva le nts of tri ethyl amine were used, and the content It was heated to 85 °C as a solution in oxygen/water (3:1) for 3 h. Ethyl acetate was used as a solvent during preparation. The organic layers were dried, filtered and concentrated under vacuum by proportioning the desired compound, the reference compound I- 390 (200 mg, yield 75%) as a white solid.

1H NMR (500 MHz, DMSO-da) 8 ppm 9.13 (s, 1 H), 8.30 (d, 1 H), 7.71 (s, 1 H), 7.31 - 7.37 ( m, 1 H), 7.19 - 7.26 (m, 2 H), 7.11 (t, 1 H), 6.98 (t, 1 H), 6.86 (t, 1 H), 5.92 (s, 2 H), 3.50-3.59 (m, 3 H), 3.25 (q, 2 H), 1.31 -1.37 (m, 9 H). Reference compound I-391

A mixture of reference compound I-390 (1 equiv.) and HCl [4.0 M in 1,4-dioxane] (50 equiv.) was stirred at 23 °C for 24 h. The mixture was concentrated in vacuo to provide the desired compound, reference compound I-391 (as HCl salt, 157 mg, 100% yield) as a white solid.

1H NMR (500 MHz, DMSO-d6) 8 ppm 9.11 (s, 1 H), 8.31 (br. s., 1 H), 7.73 (br. s., 1 H), 7, 26 - 7.38 (m, 2 H), 7.21 - 7.26 (m, 1 H), 7.11 (t, 1 H), 6.83 (t, 1 H), 5.91 ( s, 2 H), 3.74 (br. s., 2 H), 3.07-3.15 (m, 2 H).

Reference compound I-392 A mixture containing reference compound I-391 (1 equiv.), DBU (2.0 equiv.), triethylamine (2 equiv.) and N-phenyl-bis(trifluoromethanesulfonimide) (1. 2 equiv.) in acetonitrile was stirred at 23 °C for 24 h. The mixture was concentrated in vacuo. The resulting residue was diluted in ethyl acetate and washed with water. The organic layer was dried, filtered and evaporated to give an oil. The oil was purified by silica gel chromatography (gradient from 0 to 100% ethyl acetate in hexanes). Recrystallization of the purified material provided the desired compound, reference compound I-392 (123 mg, 19% yield) as a white solid.

1H NMR (500 MHz, DMSO-d6) 8 ppm 9.56 (s, 1 H), 9.11 (d, 1 H), 8.22 (d, 1 H), 7.87 (t, 1 H) ), 7.51 (s, 1 H), 7.30 - 7.35 (m, 1 H), 7.20 - 7.24 (m, 1 H), 7.08 - 7.13 (m, 2 H), 6.83 - 6.87 (m, 1 H), 5.89 (s, 2 H), 3.60 (q, 2 H), 3.46 (d, 2 H). Reference compound I-393

The title compound was synthesized in 3 steps:

Step 1: Synthesis of methyl 2-hydroxy-2-methyl-3-(methylamino)propanoate

A mixture of methanamine [2.0 M in THF] (1.3 equiv) and methyl 2-methylglycidate (1 equiv) in ethanol was heated to 80 °C for 24 h. The mixture was concentrated to give methyl 2-hydroxy-2-methyl-3-(methylamino)propanoate (2.26 g) as a clear oil. The mixture was collected in the next step without further purification.

Stage 2: Synthesis of 2-hydroxy-2-methyl-3-(methylamino)propanamide

A mixture of methyl 2-hydroxy-2-methyl-3-(methylamino)propanoate (1 equiv.) and ammonia [7.0 M in methanol] (5 equiv.) was heated to 85 °C in a sealed vial for 24 hours The mixture was concentrated to give 2-hydroxy-2-methyl-3-(methylamino)propanamide as a thick oil. The material was used in the following reaction without further purification. Stage 3: Synthesis of reference compound I-393

The title compound was prepared following general procedure B, except that 2-hydroxy-2-methyl-3-(methylamino)propanamide (4 equiv.) was the amine reagent, 4 equivalents of triethylamine were used, and the content was heated to 85 °C as a solution in dioxane/water (3:1) for 24 h. The mixture was cooled to 23°C and the white precipitate formed was collected by filtration, clarified with diethyl ether and dried under vacuum to provide the desired compound, reference compound I-393 (45 mg, 11% yield during step 3) as a white solid. 1H NMR (500 MHz, DMSO-d6) 8 ppm 9.09 (d, 1 H), 8.21 (d, 1 H), 7.52 (s, 1 H), 7.27 - 7.36 ( m, 2 H), 7.19 - 7.26 (m, 1 H), 7.10 (t, 1 H), 6.88 (t, 1 H), 5.88 (s, 2 H), 4.11 (d, 1 H), 3.72 (d, 1 H), 3.26 (d, 3 H), 1.25 (s, 3 H).

Reference compound I-394

The title compound was synthesized in 3 steps:

Step 1: Synthesis of 3-((5-fluoro-2-(1-(2-fluorobenzyl)-5-(isoxazol-3-yl)-1H-pyrazol-3-yl)pyrimidin-4-yl)amino) fed-butyl azetidine-1-carboxylate

The intermediate product was prepared by following the general procedure B, except that the 3 -a min or a ze tid in -1 - Fe d -butyl carboxylate (4 eq u iv .) was the amine reagent, 4 eq uiva le nts of tri tyla mine were used, and the con The dye was heated to 80 °C as a solution in dioxane/water (3:1) for 24 h. Ethyl acetate was used as a solvent during preparation. The organic layer was dried, filtered and evaporated giving 3-((5-fluoro-2-(1-(2-fluorobenzyl)-5-(isoxazol-3-yl)-1H-pyrazol-3-yl)pyrimidin- Fed-butyl 4-yl)amino)azetidine-1-carboxylate (505 mg, 100% yield) as a white solid.

1H NMR (500 MHz, DMSO-da) 8 ppm 9.10 (d, 1 H), 8.45 (br. s., 1 H), 8.36 (d, 1 H), 8.27 (d , 1 H), 7.53 - 7.57 (m, 1 H), 7.30-7.37 (m, 1 H), 7.19-7.28 (m, 1 H), 7.11 (t, 1 H), 6.83-6.91 (m, 1 H), 5.90 (s, 2 H), 4.81 -4.90 (m , 1 H), 4.21 (br .s., 2 H), 3.92 (dd, 2 H), 1.36 -1.41 (m, 9 H).

Stage 2: Synthesis of A/-(azet¡din-3-¡l)-5-fluoro-2-(1-(2-fluorobenzyl)-5-(¡soxazol-3-¡l)- hydrochloride 1H-pyrazol-3-yl)pyrimidin-4-amine

A mixture of 3-((5-fluoro-2-(1-(2-fluorobenzyl)-5-(isoxazol-3-yl)-1H-pyrazol-3-yl)pyrimidin-4-yl)amino)azetidin- Fed-butyl 1-carboxylate (1 equiv.) and hydrogen chloride [4.0 M in 1,4-dioxane] (10 equiv.) were stirred at 23 °C for 4 h. The mixture was concentrated in vacuo to give A/-(azetidin-3-yl)-5-fluoro-2-(1-(2-fluorobenzyl)5-(isoxazol-3-yl)-1H-pyrazol-3-yl) pyrimidine-4-amine (as HCl salt, 450 mg, 100% yield) as a white solid.

1H NMR (500 MHz, DMSO-cfe) 8 ppm 9.13 - 9.16 (m, 1 H), 8.40 - 8.49 (m, 1 H), 7.62 - 7.70 (m, 1H), 7.32 - 7.39 (m, 1H), 7.28 (dd, 1H), 7.21 - 7.25 (m, 1H), 7.12 (td, 1H ), 6.89 (t, 1 H), 5.93 (s, 2 H), 4.95 - 5.02 (m, 1 H), 4.26 - 4.35 (m, 3 H), 4.17-4.25 (m, 2 H).

Stage 3: Synthesis of reference compound I-394

To a cold mixture of A/-(azetidin-3-yl)-5-fluoro-2-(1-(2-fluorobenzyl)-5-(isoxazol-3-yl)-1H-pyrazol-3-yl hydrochloride )pyrimidin-4-amine (1 equiv.), triethylamine (3 equiv.) and pyridine (245 pl, 3 equiv.) in dichloromethane at -78 ° C trifluoromethanesulfonic anhydride (4 equiv.) was added. The mixture was stirred at −78 °C for 2 h before heating to 23 °C. The mixture was then treated with 1N HCl solution and extracted with ethyl acetate. The organic layer was dried, filtered and evaporated to give an oil. The oil was purified by silica gel chromatography (gradient from 0 to 30% ethyl acetate in hexanes) to provide the desired compound, reference compound I-394 (90 mg, 16% yield) as a white solid.

1H NMR (500 MHz, CD3OD) 8 ppm 8.83 (d, 1 H), 8.32 (d, 1 H), 7.62 (s, 1 H), 7.29 - 7.34 (m, 1 H), 7.05 - 7.13 (m, 2 H), 6.96 - 7.00 (m, 2 H), 6.03 (s, 2 H), 4.98 (br. s. , 2 H), 4.74 - 4.83 (m, 1 H), 4.53 (br. s., 2 H).

Reference compound I-395

The title compound was prepared following general procedure B, except that (1R,2S)-(+)-c/'s-1-amino-2-indanol (4 equiv.) was the amine reagent, 4 equivalents of triethylamine, and the content was heated to 80 °C as a solution in dioxane/water (3:1) for 3 h. Ethyl acetate was used as a solvent during preparation. The crude material was purified by silica gel chromatography with a gradient of 0-50% ethyl acetate/hexanes providing the desired compound, reference compound I-395 (4 mg, 27% yield) as a cream-colored solid. .

1H NMR (500 MHz, DMSO-d6) 8 ppm 9.07 (d, 1 H), 8.28 (d, 1 H), 7.55 (s, 1 H), 7.30 - 7.37 ( m, 2 H), 7.26 - 7.29 (m, 1 H), 7.13 - 7.25 (m, 5 H), 7.10 (td, 1 H), 6.80 - 6, 87 (m, 1 H), 5.89 (s, 2 H), 5.75 (dd, 1 H), 5.27 (d, 1 H), 4.60 (d, 1 H), 3, 15 (dd, 1 H), 2.93 (dd, 1 H).

Reference compound I-396

The title compound was prepared following general procedure B, except that 3-(trifluoromethyl)azetidin-3-ol (3 equiv.) was the amine reagent, 3 equivalents of triethylamine were used, and the contents were heated to 80 °C as a solution in dioxane/water (3:1) for 3 h. Ethyl acetate was used as a solvent during preparation. The organic layer was dried, filtered and concentrated in vacuo to give a solid, which was rinsed with minimal amounts of methanol and diethyl ether and dried in vacuo to give the desired compound, reference compound I-396 (57 mg, yield 45%) as a white solid.

1H NMR (500 MHz, DMSO-d6) 8 ppm 9.09 (d, 1 H), 8.37 (d, 1 H), 7.57 (s, 2 H), 7.30 - 7.36 ( m, 1 H), 7.19 - 7.26 (m, 2 H), 7.10 (td, 1 H), 6.82 (t, 1 H), 5.91 (s, 2 H), 4.52 (d, 2 H), 4.30 (d, 2 H).

Reference compound I-397

The title compound was prepared following general procedure B, except that 4-(aminomethyl)tetrahydro-2H-pyran-4-ol (3 equiv.) was the amine reagent, 3 equivalents of triethylamine were used, and the content was heated to 80 °C as a solution in dioxane/water (3:1) for 24 h. Ethyl acetate was used as a solvent during preparation. The organic layer was dried, filtered and concentrated in vacuo to provide the desired compound, reference compound I-397 (30 mg, 24% yield) as a white solid.1

1H NMR (500 MHz, CD3OD) 8 ppm 8.80 (d, 1 H), 8.20 (d, 1 H), 7.57 (s, 1 H), 7.24 - 7.32 (m, 1 H), 7.02 - 7.13 (m, 2 H), 6.87 - 6.97 (m, 2 H), 5.99 (s, 2 H), 3.74 - 3.80 ( m, 6 H), 1.76 - 1.85 (m, 2 H), 1.59 (d, 2 H).

I-398 REFERENCE COMPOSITION

The title compound was prepared following general procedure B, except that 2-amino-4,4-difluorobutanoic acid (3 equiv.) was the amine reagent, 3 equivalents of triethylamine were used, and the contents were heated to 80 °C as a solution in dioxane/water (4:1) for 24 h. Ethyl acetate was used as a solvent during the work. The organic layer was dried, filtered and concentrated in vacuo to give a solid, which was rinsed with minimal amounts of methanol and diethyl ether and dried in vacuo to give the desired compound, reference compound I-398 (123 mg, yield 52%) as a white solid.

1H NMR (500 MHz, DMSO-da) 8 ppm 9.10 (d, 1 H), 8.29 (d, 1 H), 8.12 (d, 1 H), 7.42 (s, 1 H) ), 7.32 (d, 1 H), 7.18 - 7.24 (m, 1 H), 7.16 (d, 1 H), 7.10 (t, 1 H), 6.85 ( t, 1 H), 6.09 - 6.37 (m, 2 H), 5.87 (s, 2 H), 4.85 (d, 1 H), 4.04 (s, 1 H).

Reference compound I-399

To a suspension of the reference compound I-398 (1 equiv.) in THF was added lithium aluminum hydride [1.0 M solution in THF] (3 equiv.). The mixture was stirred at 23 °C for a further 24 h. The mixture was quenched with water, then a 15% NaOH solution, then water. The mixture was diluted with ethyl acetate and washed with 1N HCl solution. The organic layer was dried, filtered and evaporated to give an oil. The oil was purified by silica gel chromatography (gradient from 0 to 100% ethyl acetate in hexanes) to provide the desired compound, reference compound I-399 (23 mg, 12% yield) as a white solid.

1H NMR (500 MHz, CD3OD) 8 ppm 8.75 (d, 1 H), 8.08 (d, 1 H), 7.43 (s, 1 H), 7.23 - 7.30 (m, 1 H), 7.06 - 7.12 (m, 1 H), 7.03 (t, 1 H), 6.87 - 6.89 (m, 1 H), 6.81 (t, 1 H) ), 6.05 - 6.21 (m, 1 H), 5.93 - 5.98 (m, 2 H), 4.71 (dq, 1 H), 3.63 - 3.75 (m, 2H), 2.18-2.35 (m, 2H).

Reference compound I-400

A mixture of 2,2 bis-(trifluoromethyl)-2-hydroxyacetic acid (1.5 equiv.) and CDI (1.5 equiv.) in THF was refluxed for 2 h. To this mixture was added 2-(1-(2-fluorobenzyl)-5-(isoxazol-3-yl)-1H-pyrazol-3-yl)pyrimidin-4-amine (previous intermediate described in the application publication patent WO2012/3405 A1) (1 equiv.) in one part. The mixture was heated at reflux for 3 h. It was then cooled to 23 °C, diluted in ethyl acetate and washed with 1N HCl solution. The organic layer was dried, filtered and evaporated to give an oil. The oil was purified by silica gel chromatography (gradient from 0 to 80% ethyl acetate in hexanes) to provide the desired compound, reference compound I-400 (111 mg, 35% yield) as a white solid.

1H NMR (500 MHz, CD3OD) 8 ppm 8.79 - 8.82 (m, 1 H), 8.75 - 8.77 (m, 1 H), 8.07 - 8.10 (m, 1 H) ), 7.54 - 7.56 (m, 1 H), 7.22 - 7.31 (m, 1 H), 6.99 - 7.12 (m, 2 H), 6.86 - 6, 93 (m, 2 H), 5.95 - 6.00 (m, 2 H).

Reference compound I-401

The title compound was synthesized in 2 steps:

Step 1: Synthesis of (S)-4-((5-fluoro-2-(1-(2-fluorobenzyl)-5-(isoxazol-3-yl)-1H-pyrazol-3-yl)pyrimidin-4 acid -yl)amino)-5-methoxy-5-oxopentanoic acid

The intermediate product was prepared following general procedure B, except that L-glutamate methyl ester (4 equiv.) was the amine reagent, 4 equivalents of triethylamine were used, and the contents were heated to 80 °C as a solution. in dioxane/water (3:1) for 24 h. The mixture was concentrated in vacuo. The resulting solid was clarified with diethyl ether and water, and collected by filtration and dried under vacuum to provide acid-(isoxazol-3-yl)-1H-pyrazol-3-yl)pyrimidin-4-yl)amino)-5 -methoxy-5-oxopentanoic acid (274 mg, 69% yield) as a yellow solid.12

1H NMR (500 MHz, CD3OD) 8 ppm 8.76 - 8.78 (m, 1 H) 8.15 (d, 1 H) 7.37 (s, 1 H) 7.23 - 7.31 (m , 2 H) 6.99 - 7.13 (m, 2 H) 6.84 (t, 1 H) 5.94 - 5.99 (m, 2 H) 3.73 (s, 1 H) 3. 34 (s, 3 H) 2.29 - 2.43 (m, 2 H) 2.12 - 2.24 (m, 2 H).

Stage 2: Synthesis of reference compound I-401

To a solution of acid (S)-4-((5-fluoro-2-(1-(2-fluorobenzyl)-5-(isoxazol-3-yl)-1H-pyrazol-3-yl)pyrimidin-4- yl)amino)-5-methoxy-5-oxopentanoic acid (1 equiv.) in THF, oxalyl chloride (1.5 equiv.) and a drop of DMF were added. The mixture was stirred at 23 °C for 30 min. It was then heated at reflux for 30 min. The mixture was cooled to 23 °C, diluted with ethyl acetate and washed with 1N HCl solution. The organic layer was dried, filtered and evaporated to give an oil. The oil was purified by silica gel chromatography (gradient from 0 to 50% ethyl acetate in hexanes) to provide the desired compound, reference compound I-401 (81 mg, 33% yield) as a light yellow solid.

1<H R M N (500 M H z, C D 3O D ) 8 ppm 8 .76 - 8 .78 (m, 1 H), 8.71 - 8 .74 (m, 1 H), 7 .32 - 7 .35 ( m, 1 H), 7 .23 - 7 .30 (m, 1 H),>7 .06 - 7 .12 (m, 1 H), 7 .00 - 7 .05 (m, 1 H), 6 .81 - 6 .90 (m, 2 H), 5 .94 (s, 2 H), 5 .10 (dd, 1 H), 3 .75 (s, 3 H), 2 .59 - 2 .80 (m, 3 H), 2 .23 - 2 .33 (m , 1 H).

I-403 REFERENCE COMPOSITION

The title compound was prepared following general procedure B, except that 2-amino-3,3,3-trifluoropropanoic acid (3 equiv.) was the amine reagent, 3 equivalents of triethylamine were used, and the content was heated at 70 °C for 24 h as a solution in dioxane/water (3:1). Ethyl acetate was used as a solvent during preparation. The organic layer was dried, filtered and concentrated in vacuo to give a solid, which was rinsed with minimal amounts of methanol and diethyl ether and dried in vacuo to give the desired compound, reference compound I-403 (71 mg, yield 17%) as a yellow solid.

1H NMR (500 MHz, CD3OD) 8 ppm 8.76 (d, 1 H), 8.27 (d, 1 H), 7.48 - 7.51 (m, 1 H), 7.24 - 7, 31 (m, 1 H), 7.07 - 7.13 (m, 1 H), 7.00 - 7.06 (m, 1 H), 6.92 (d, 1 H), 6.80 ( t, 1 H), 6.10 - 6.16 (m, 1 H), 5.98 (s, 2 H).

Reference compound I-404

To a suspension of the reference compound I-403 (1 equiv.) in THF was added lithium aluminum hydride [1.0 M solution in THF] (2 equiv.). The mixture was stirred at 23 °C for 1 h. The mixture was quenched with water, then a 15% NaOH solution, then water. The mixture was diluted with ethyl acetate and washed with 1N HCl solution. The organic layer was dried, filtered and evaporated to give a solid. The solid was purified by silica gel chromatography (gradient from 0 to 50% ethyl acetate in hexanes) to provide the desired compound, reference compound I-404 (19 mg, 12% yield) as a white solid.

1H NMR (500 MHz, CD3OD) 8 ppm 8.75 - 8.77 (m, 1 H), 8.21 (d, 1 H), 7.46 - 7.48 (m, 1 H), 7, 24 - 7.31 (m, 1 H), 7.06 -7.14 (m, 1 H), 7.03 (t, 1 H), 6.89 - 6.94 (m, 1 H), 6.79 (t, 1 H), 5.97 (d, 2 H), 5.47 - 5.54 (m, 1 H), 3.87 - 4.01 (m, 2 H).

Reference compound I-405

The title compound was synthesized in 3 steps:

Stage 1: Synthesis of 2-(trifluoromethyl)oxirane-2-carboxamide

To a solution of 2-(bromomethyl)-3,3,3-trifluoro-2-hydroxypropanamide (1 equiv.) in acetone was added potassium carbonate (2 equiv.). The mixture was heated at reflux for 2 h. The mixture was concentrated in vacuo. The resulting residue was diluted with water and extracted with ethyl acetate. The organic layer was dried, filtered and evaporated to give 2(trifluoromethyl)oxirane-2-carboxamide (1.44 g, 76% yield) as a yellow gum.

1H NMR (500 MHz, CD3OD) 8 ppm 3.17 (dd, 2 H).

Stage 2: Synthesis of 2-(aminomethyl)-3,3,3-trifluoro-2-hydroxypropanamide

A mixture of ammonia [7 M in methanol] (10 equiv.) and 2-(trifluoromethyl)oxirane-2-carboxamide (1 equiv.) was stirred in a sealed vial at 80 °C for 24 h. The mixture was concentrated in vacuo to provide 2-(aminomethyl)-3,3,3-trifluoro-2-hydroxypropanamide (1.3 g, 84% yield) as a brown gum.

1H NMR (500 MHz, DMSO-d6) 83.01 - 3.11 (m, 1 H), 2.84 (d, 1 H).

Stage 3: Synthesis of reference compound I-405

The title compound was prepared following general procedure B, except that 2-(aminomethyl)-3,3,3-trifluoro-2-hydroxypropanamide (4 equiv.) was the amine reagent, 4 equivalents of triethylamine were used, and the content was heated at 90 °C for 24 h as a solution in dioxane/water (3:1). The mixture was diluted in ethyl acetate and washed with water. The organic layer was dried, filtered and evaporated to give a solid. The solid was purified by silica gel chromatography (gradient from 0 to 80% ethyl acetate in hexanes) to provide the desired compound, reference compound I-405 (262 mg, 40% yield) as a white solid.

1H NMR (500 MHz, DMSO-cfe) 8 ppm 9.08 - 9.13 (m, 1 H), 8.33 (d, 1 H), 7.49 - 7.55 (m, 1 H), 7.28 - 7.37 (m, 1 H), 7.17 - 7.25 (m, 2 H), 7.10 (t, 1 H), 6.98 (t, 1 H), 5, 86 - 5.92 (m, 2 H), 3.92 - 4.04 (m, 2 H).

Reference compound I-406

The title compound was prepared by following the general procedure B, except that the acid 3 -methylp ip e rid in -3 -c a rb o x ylic (as HCl salt, 1.7 eq u iv .) was the amine reagent, and the cont ent was heated to 100 °C for 19 h.

The reaction mixture was acidified to pH 3 with a 1N HCl solution and the resulting solids were collected by vacuum filtration. providing the desired compound, the reference compound I-406 (95 mg, yield 94%) as a white solid.

1H-NMR (500 MHz, CD3OD) 88.75 (d, 1 H), 8.10 (d, 1 H), 7.40 (s, 1 H), 7.26 (ap. q, 1 H) , 7.09 (dd, 1 H), 7.03 (ap. t, 1 H), 6.91 (d, 1 H), 6.81 (ap. t, 1 H), 5.95 (s , 2 H), 4.58 (d, 1 H), 4.39 (br d, 1 H), 3.43 (m, 1 H), 3.41 (d, 1 H), 2.24 ( m, 1 H), 1.77 (m, 2 H), 1.61 (m, 1 H), 1.23 (s, 3 H).

Reference compound I-407

The title compound was prepared following general procedure B, except that 2-aminoethanol (10 equiv.) was the amine reagent, no triethylamine was used, and the contents were heated at 100 °C for 40 min. The reaction mixture was acidified to pH 3 with 1N HCl solution and the resulting solids were collected by vacuum filtration to provide the desired compound, reference compound I-407 (57 mg, 58% yield) as a white solid.

1H-NMR (500 MHz, DMSO-d6) 89.08 (d, 1 H), 8.17 (d, 1 H), 7.61 (ap. t, 1 H), 7.50 (s, 1 H), 7.33 (ap. q, 1 1 H), 7.22 (ap. t, 1 H), 7.10 (ap. t, 1 H), 6.82 (ap. t, 1 H ), 5.89 (s, 2 H), 4.78 (t, 1 H), 3.58 (m, 4 H).

Reference compound I-408

The title compound was prepared following general procedure B, except that 3-aminopropan-1-ol (10 equiv.) was the amine reagent, no triethylamine was used, and the contents were heated at 100 °C for 2 h. The reaction mixture was acidified to pH 3 with 1N HCl solution and the resulting solids were collected by vacuum filtration to provide the desired compound, reference compound I-408 (26 mg, 37% yield) as an off-white solid.

1H-NMR (500 MHz, DMSO-d6) 89.08 (d, 1 H), 8.16 (d, 1 H), 7.66 (ap. t, 1 H), 7.49 (s, 1 H), 7.32 (ap. q, 1 1 H), 7.21 (m, 1 H), 7.10 (ap. t, 1 H), 6.83 (ap. t, 1 H), 5.89 (s, 2 H), 4.54 (t, 1 H), 3.51 (m, 4 H), 1.75 (ap. quintuplet, 2 H).

Reference compound I-409

The title compound was synthesized in 2 steps:

Stage 1. Synthesis of 1-(((terf-butoxycarbonyl)(ethyl)amino)methyl)cyclopropanecarboxylic acid

A solution of 1-(((tert-butoxycarbonyl)amino)methyl)cyclopropanecarboxylic acid in THF at 0 °C was treated with iodoethane (10 equiv.) followed by sodium hydride (mineral oil 60% w/w, 10 equivalents, added in 6 parts).

After 2 days, the reaction mixture was carefully quenched with water and washed with ethyl acetate. The aqueous layer was acidified to pH 3 with 3N HCl and extracted with ethyl acetate. The organic phases were dried over sodium sulfate, filtered and the solvent was removed in vacuo providing 1-(((tert-butoxycarbonyl)(ethyl)amino)methyl)cyclopropanecarboxylic acid (92% yield) as a pale yellow oil which was used. without additional purification.

Stage 2. Synthesis of A/-((1-carboxycyclopropyl)methyl)ethanaminium trifluoroacetate

A solution of 1-(((tert-butoxycarbonyl)(ethyl)amino)methyl)cyclopropanecarboxylic acid in dichloromethane was treated with trifluoacetic acid (30 equiv.). After 2 h, the reaction mixture was concentrated in vacuo to provide W-((1-carboxycyclopropyl)methyl)ethanaminium trifluoroacetate (yield >99%) as a dark yellow oil which was used without further handling.

Stage 3: Synthesis of reference compound I-409

The title compound was prepared following general procedure B, except that N-((1-carboxycyclopropyl)methyl)ethanaminium trifluoroacetate (1.5 equiv.) was the amine reagent, and the contents were heated to 100 °C for 3 days. The crude material was purified by reverse phase HPLC (20-70% acetonitrile/water gradient with 0.1% TFA) providing the desired product, reference compound I-409 (86 mg, 51% yield) as a white solid. 1H-NMR (500 MHz, CD3OD) 88.82 (d, 1 H), 8.28 (d, 1 H), 7.52 (s, 1 H), 7.30 (ap. q, 1 H) , 7.10 (m, 1 H), 7.06 (ap. t, 1 H), 6.97 (d, 1 H), 6.96 (m, 1 H), 6.00 (s, 2 H), 4.30 (s, 2 H), 4.00 (q, 2 H), 1.41 (q, 2 H), 1.37 (t, 3 H), 1.16 (q, 2 H).

I-410 REFERENCE COMPOSITION

The title compound was synthesized in 5 steps:

Step 1: Synthesis of ethyl 1-(2-fluorobenzyl)-5-methyl-1H-pyrazole-3-carboxylate

To ethyl 3-methyl-1H-pyrazole-5-carboxylate in DMF was added sodium hydride (60% by weight in mineral oil, 1.2 equiv.). After 10 min, 2-fluorobenzyl bromide (1.2 equiv.) was added and the reaction was stirred for 20 h. Water was added and the resulting mixture was extracted with ethyl acetate. The combined organic phases were washed with water and brine, dried over sodium sulfate, filtered, and the solvent was removed in vacuo. Purification by silica gel chromatography (10-40% ethyl acetate/hexanes gradient) afforded ethyl 1-(2-fluorobenzyl)-5-methyl-1H-pyrazole-3-carboxylate (79% yield) and 1 -(2-fluorobenzyl)-3-methyl-1H-pyrazole-5-carboxylate ethyl (yield 9%).

Stage 2: Synthesis of 1-(2-fluorobenzyl)-5-methyl-1H-pyrazole-3-carboxylic acid

To a solution of ethyl 1-(2-fluorobenzyl)-5-methyl-1H-pyrazole-3-carboxylate in THF/MeOH/water (3:1:1 ratio) was added lithium hydroxide hydrate (1.5 equiv .). After 23 h, the volatile organic materials were removed in vacuo and the resulting mixture was acidified to pH 3 with 1N HCl. 1-(2-fluorobenzyl)-5-methyl-1H-pyrazole-3-carboxylic acid was collected by vacuum filtration (92% yield).

Step 3: Synthesis of 1-(2-fluorobenzyl)-5-methyl-1H-pyrazole-3-carbonitrile

To a suspension of 1-(2-fluorobenzyl)-5-methyl-1H-pyrazole-3-carboxylic acid, 2-methylpropan-2-amine (3 equiv.), and triethylamine (2 equiv.) in ethyl acetate, added W-propylphosphonic anhydride (T3P, 50% solution in ethyl acetate, 3 equiv.). The resulting yellow solution was heated at 65 °C for 2.5 h. The solvent was removed in vacuo. Phosphoryl trichloride (12 equiv.) was added and the resulting mixture was stirred at 70 °C for 1 h 40 min. The reaction was quenched by carefully pouring into a mixture of ice and water, neutralized to pH 7 by the addition of saturated sodium bicarbonate solution, and extracted with dichloromethane. The combined organic phases were dried over sodium sulfate, filtered, and the solvent was removed in vacuo. Purification by silica gel chromatography (10% ethyl acetate/hexanes gradient) afforded 1-(2-fluorobenzyl)-5-methyl-1H-pyrazole-3-carbonitrile (49% yield).

Step 4: Synthesis of 1-(2-fluorobenzyl)-5-methyl-1H-pyrazole-3-carboxymide

A solution of 1-(2-fluorobenzyl)-5-methyl-1H-pyrazole-3-carbonate in methanol was treated with sodium methoxide (solution to 25% by weight in MeOH, 5 equiv.) and stirred for 24 h. Ammonium chloride (10 equiv.) was added. After 26 h, the reaction mixture was concentrated in vacuo and partitioned between semisaturated sodium bicarbonate and ethyl acetate. The combined organic phases were dried over sodium sulfate, filtered, and the solvent was removed in vacuo. The raw product was contaminated with starting material due to the incomplete reaction. This material was subjected to similar conditions providing 1-(2-fluorobenzyl)-5-methyl-1H-pyrazole-3-carboxy dam¡da (yield 92 %). Stage 5: Syntheses of the reference compound I-410

A suspension of 1-(2-fluorobenzyl)-5-methyl-1H-pyrazol-3-carboxymide was treated with (Z)-3-ethoxy-2- sodium fluoro-3-oxoprop-1-en-1-olate (see also general procedure A, step 4, 3.0 equiv.) and heated to 90 ° C for 1 hour. After cooling to room temperature, the reaction mixture was neutralized by the addition of HCl (1.25 M solution in EtOH). The resulting tan suspension was concentrated in vacuo. The residue was partitioned between dichloromethane and water and the aqueous layer was extracted again with dichloromethane. The combined organic phases were dried over sodium sulfate, filtered, and the solvent was removed in vacuo. Trituration with dichloromethane afforded the title compound (206 mg, yield 62%) as a white solid.

1H-NMR (500 MHz, DMSO-da) 812.9 (brs, 1 H), 8.07 (brs, 1 H), 7.38 (ap. q, 1 H), 7.25 (m, 1 H), 7.18 (ap. t, 1 H), 7.11 (m, 1 H), 6.72 (s, 1 H), 5.44 (s, 2 H), 2.30 (s , 3H).

Reference compound I-411

The title compound was synthesized in 2 steps:

Stage 1: Syntheses of 4-chloro-5-fluoro-2-(1-(2-fluorobenzyl)-5-methyl-1H-pyrazol-3-l) pyrm dna

The intermediate product was generated using the procedure described for the synthesis of intermediate product 1, except for the use of the reference compound I-410 as the parent. my starting point. An off-white solid was obtained (210 mg, yield 96%), and the content was used without further purification.

Stage 2: Syntheses of the reference compound I-411

The title compound was prepared following general procedure B, except that 1-((methylamino)methyl)cyclopropanecarboxylic acid (as the salt of HCl) was the amine reactant. The reaction mixture was acidified to pH 3 and the resulting precipitate was collected by vacuum filtration providing the desired compound, the reference compound. I-411 (48 mg, yield 93%) as a white solid.

1H-NMR (500 MHz, DMSO-d6) 812.3 (s, 1 H), 8.16 (d, 1 H), 7.36 (ap. q, 1 H), 7.25 (m, 1 H), 7.17 (ap. t, 1 H), 6.99 (ap. t, 1 H), 6.61 (s, 1 H), 5.38 (s, 2 H), 3.97 (s, 2 H), 3.22 (d, 3 H), 2.29 (s, 3 H), 1.13 (m, 2 H), 1.01 (m, 2 H).

Reference compound I-412

The title compound was prepared following general procedure B, except that (2R,3S)-3-methylperidin-2-carboxylic acid was the amine reagent, and the contents were heated at 100 °C for 20 h. The crude material was purified by reverse phase HPLC (gradient of 25-80% acetonitril/water with 0.1% TFA) providing the desired compound, the reference compound I-412 (12 mg, yield 23%) as a white solid.

1H-NMR (500 MHz, CD3OD) 88.33 (d, 1 H), 7.36 (ap. q, 1 H), 7.15 (m, 2 H), 7.06 (ap. t, 1 H), 6.89 (s, 1 H), 5.52 (s, 2 H), 5.36 (d, 1 H), 4.58 (br s, 1 H), 3.83 (ap. t, 1 H), 2.36 (s, 3 H), 1.99 (m, 1 H), 1.96 (m, 1 H), 1.79 (m, 2 H), 1.58 ( m, 1 H), 1.24 (d, 3 H).

Reference compound I-413

The title compound was prepared following general procedure B, except that sernol (10 equiv.) was the amine reagent, and no triet was used. foil, and the contents were heated to 90 °C for 40 min and then to 100 °C for 20 min. The reaction mixture was acidified to pH 3 and the resulting precipitate was collected by vacuum filtration providing the desired compound, the reference compound. I-413 (46 mg, yield 88%) as a white solid.

1H-NMR (500 MHz, DMSO-d6) 88.11 (d, 1 H), 7.35 (ap. q, 1 H), 7.24 (m, 1 H), 7.16 (ap. t , 1 H), 7.03 (d, 1 H), 6.92 (ap. t, 1 H), 6.64 (s, 1 H), 5.40 (s, 2 H), 4.71 (t, 2 H), 4.24 (m, 1 H), 3.56 (m, 4 H), 3.12 (s, 3 H).

Reference compound I-414

The title compound was prepared following general procedure B, except that 2 -(a minimum)-1,1,1, 3 ,3 ,3 -h e xa flu or ro prop a n -2 -o l (2 ,2 equiv.) was the amine reagent, 5 eq uval le ns of three were used etilamine and the content was heated to 90 °C for 18 h and then to 100 °C for 4 days. The crude material was purified by reverse phase HPLC (gradient of 35-80% acetonitrile/water with 0.1% TFA) providing the desired compound, reference compound I -414 (35 mg, 58% yield) as a white solid.

1H-NMR (500 MHz, MeOH-d4) 88.29 (d, 1 H), 7.35 (ap. q, 1 H), 7.13 (m, 3 H), 6.79 (s, 1 H), 5.50 (s, 2 H), 4.29 (s, 2 H), 2.36 (s, 3 H).

Reference compound I-416

The title compound was synthesized in 3 steps:

Stage 1: Synthesis of 3-(1-(2-fluorobenzyl)-5-(isoxazol-3-yl)-1H-pyrazol-3-yl)-1,2,4-triazin-5(4H)-one.

A solution of 1-(2-fluorobenzyl)-5-(isoxazol-3-yl)-1H-pyrazole-3-carboximidamide hydrochloride in absolute ethanol was treated with hydrazine hydrate (1.5 equiv.) at room temperature. After 45 min, ethyl 2-oxoacetate (50 wt % solution in toluene, 3.0 equiv.) was added and the resulting solution was heated at 50-60 °C for 65 h. The reaction mixture was then concentrated in vacuo, taken up in dichloromethane, filtered and concentrated in vacuo to give an orange oil. Purification by silica gel chromatography (10-20% acetonitrile-methanol (7:1) in dichloromethane) gave the desired compound 3-(1-(2-fluorobenzyl)-5-(isoxazol-3-yl)-1H -pyrazol-3-yl)-1,2,4-triazin-5(4H)-one (320 mg, 73% yield) as a light tan solid.

1H-NMR (500 MHz, CDCla) 88.54 (d, 1 H), 7.84 (br s, 1 H), 7.44 (s, 1 H), 7.31 (m, 1 H), 7.12-7.04 (m, 3 H), 6.63 (d, 1 H), 5.95 (s, 2 H).

Step 2: Synthesis of 3-(3-(5-chloro-1,2,4-triazin-3-yl)-1-(2-fluorobenzyl)-1H-pyrazol-5-yl)isoxazole

3-(1-(2-fluorobenzyl)-5-(isoxazol-3-yl)-1H-pyrazol-3-yl)-1,2,4-triazin-5(4H)-one was treated with phosphoryl trichloride (42 equivalents, excess). The resulting mixture was heated at 65 °C for 4 h. The resulting tan suspension was blow dried in a stream of nitrogen and azeotropically dried with toluene. The chloro-triazine was used in the next step without additional manipulation.

Stage 3: Synthesis of reference compound I-416

The title compound was prepared following general procedure B, except that serinol (10 equiv.) was the amine reagent, 3-(3-(5-chloro-1,2,4-triazin-3-yl) was used )-1-(2-fluorobenzyl)-1H-pyrazol-5-yl)isoxazole instead of intermediate 1, no triethylamine was used, and the content was heated at 100 °C for 24 h as a solution in dioxane/DMSO (7.5:1). The reaction mixture was diluted with water, neutralized to pH 4 with 1N HCl solution and extracted with dichloromethane/isopropanol (4:1). The combined organic phases were dried over sodium sulfate, filtered, and the solvent was removed in vacuo. The crude material was purified by reverse phase HPLC (gradient 70% acetonitrile/water with 0.1% TFA) to provide the desired compound, reference compound I-416 (5.2 mg, 9.8% yield). like a white solid.

1H-NMR (500 MHz, CD3OD) 88.84 (d, 1 H), 8.35 (s, 1 H), 7.74 (s, 1 H), 7.32 (ap. q, 1 H) , 7.11 (m, 1 H), 7.08 (ap. t, 1 H), 6.99 (ap. t, 1 H), 6.96 (d, 1 H), 6.06 (s , 2 H), 4.62 (ap. quintuplet, 1 H), 3.84 (m, 4 H).

Reference compound I-417

The title compound was prepared following general procedure B, except that glycinamide (as HCl salt, 3.0 equiv.) was the amine reagent, and the contents were heated at 100 °C for 22 h. The reaction mixture was acidified to pH 4 with a 1N HCl solution and the resulting solids were collected by vacuum filtration to provide the desired compound, reference compound I-417 (110 mg, 94% yield) as a white solid.

1H-NMR (500 MHz, DMSO-d6) 89.09 (d, 1 H), 8.22 (d, 1 H), 7.83 (ap. t, 1 H), 7.51 (br s, 1 H), 7.48 (s, 1 H), 7.31 (ap. q, 1 H), 7.21 (m, 1 H), 7.18 (d, 1 H), 7.12- 7.08 (m, 2 H), 6.81 (ap. t, 1 H), 5.89 (s, 2 H), 4.01 (d, 2 H).

Reference compound I-418

The title compound was prepared following general procedure B, except that 3-amino-4,4,4-trifluorobutanoic acid (3 equiv.) was the amine reagent, and the contents were heated at 100 °C for 3. 5 hours, then at 120 °C for 18 h as a solution in dioxane/DMSO (2:1). An additional 2 equivalents of intermediate 1 was added to the reaction, and the contents were stirred at 120 °C for an additional 4 d. A mixture of dichloromethane:isopropanol (4:1) was used as a solvent during the preparation. The crude material was purified by silica gel chromatography (5% methanol/isocratic dichloromethane) to provide the desired compound, reference compound I-418 (38 mg, 10% yield) as an off-white solid.

1H NMR (500 MHz, DMSO-d6) 8 ppm 12.65 (br. s., 1 H), 9.09 (d, 1 H), 8.35 (d, 1 H), 8.29 (d , 1 H), 7.51 (s, 1 H), 7.32 (q, 1 H), 7.26 (d, 1 H), 7.24-7.19 (m, 1 H), 7 .10 (t, 1 H), 6.83 (t, 1 H), 5.91 (s, 2 H), 5.53 (br. s., 1 H), 2.96-2.85 ( m, 2H).

REFERENCE COMPOSITION I-419

A solution of reference compound I-418 in dichloromethane was treated with W,W-diisopropylethylamine (2.0 equiv.) followed by 0-(7-azabenzotriazol-1-yl)-W,W,W',W hexafluorophosphate '-tetramethyluronium (HAT, 1.2 equiv.). After 15 min, ammonia (0.5 N solution in dioxane, 2.0 equiv.) was added and the resulting light brown suspension was stirred for 1.5 h. The reaction mixture was diluted with water and extracted with dichloromethane/2-propanol (4:1). The combined organic phases were dried over magnesium sulfate, filtered, and the solvent was removed in vacuo. Purification by silica gel chromatography (0-3% methanol in dichloromethane gradient) provided the desired compound, reference compound I-419 (20 mg, 57% yield) as an off-white solid.

1H-NMR (500 MHz, DMSO-da) 89.10 (d, 1 H), 8.34 (d, 1 H), 8.24 (d, 1 H), 7.51 (m, 2 H) , 7.32 (ap. q, 1 H), 7.25 (d, 1 H), 7.22 (ap. t, 1 H), 7.10 (ap. t, 1 H), 7.06 (br s, 1 H), 6.82 (ap. t, 1 H), 5.91 (s, 2 H), 5.55 (br s, 1 H), 2.77 (dd, 1 H) , 2.67 (dd, 1 H).

Reference compound I-420

The title compound was prepared in 2 steps:

Stage 1: Synthesis of diethyl 2-(dicyanomethyl)-2-methylmalonate.

A mixture of diethyl 2-bromo-2-methylmalonate (1 equiv.), malononitrile (1 equiv.), and potassium t-butoxide (1 equiv.) in THF was heated at reflux for 15 h. The mixture was diluted with ethyl acetate and saturated aqueous ammonium chloride solution and the phases were separated. The aqueous phase was extracted twice with ethyl acetate. The combined organic phase was washed with brine, dried over anhydrous sodium sulfate, filtered and concentrated to an oil. The oil was purified by silica gel chromatography (gradient 10-15% ethyl acetate in hexane) to give 2-(dicyanomethyl)-2-methylmalonate (5.76 g, 32% yield) as a colorless oil.

1H NMR (500 MHz, CDCfe) 8 ppm4.53(s, 1 H), 4 .27-4.39 (m, 4 H), 1.81 (s, 3 H), 1.33 (t, 6 H).

Stage 2: Synthesis of reference compound I-420

A mixture of 1-(2-fluorobenzyl)-5-(isoxazol-3-yl)-1H-pyrazole-3-carboximidamide hydrochloride (generated in step 3 of general procedure A, using 1-(isoxazol-3-yl )ethanone in step 1 and 2-fluorobenzylhydrazine in step 2) (1 equiv.), diethyl 2-(dicyanomethyl)-2-methylmalonate (1.15 equiv.) and potassium bicarbonate (2 equiv.) in t -BuOH was heated at reflux for 5 h. After cooling, water was added to the reaction mixture and stirred for 30 min. The precipitate was filtered, washed with a minimum amount of water and diethyl ether and dried overnight under high vacuum to give the reference compound I-420 (385 mg, 52% yield) as a white solid.

1H NMR (500 MHz, DMSO-d6) 8 ppm 11.30 (s, 1 H), 9.10 (d, 1 H), 7.38 (s, 1 H), 7.29 - 7.36 ( m, 1 H), 7.18-7.26 (m, 2 H), 7.08-7.14 (m, 1 H), 6.81 -6.90 (m, 1 H), 6, 65 (br. s., 2 H), 5.88 (s, 2 H), 4 .04-4.16 (m, 2 H), 1.59 (s, 3 H), 1.11 (t , 3H).

Reference compound I-421

Ammonia (7.0 M in MeOH) (200 equiv.) was added to the reference compound I-420 (1 equiv.). The reaction mixture was heated at 50 °C for 16 h. The resulting solution was then concentrated in vacuo and the residue was purified by reverse phase HPLC (5-60% acetonitrile in water with 1% TFA) to provide the desired compound, reference compound I-421 (24 mg, yield 63%) as a white solid.

1H NMR (400 MHz, DMSO-cfe) 8 ppm 11.35 (br. s., 1 H), 9.08 - 9.13 (m, 1 H), 7.47 (s, 1 H), 7 .43 (s, 1 H), 7.28 - 7.38 (m, 1 H), 7.23 - 7.27 (m, 1 H), 7.17 - 7.23 (m, 2 H) , 7.06 - 7.14 (m, 1 H), 6.77 - 7.00 (m, 3 H), 5.91 (s, 2 H), 1.56 (s, 3 H).

Reference compound I-422

Cyclopropyl amine (150 equiv.) was added to the reference compound I-420 (1 equiv.) and the reaction mixture was heated at 50 °C for 30 h. The resulting solution was then concentrated in vacuo and the residue was purified by reverse phase HPLC (25-50% acetonitrile in water with 1% TFA) to provide the desired compound, reference compound I-422 (29 mg, yield 57%) as a white solid.

1H R M N (400 M Hz, DMSO-cfe) 8 ppm 11 .29 (br. s., 1 H), 9.11 (d, 1 H), 7 .59 - 7 .66 (m, 1 H), 7 .42 (s, 1 H), 7 .29 - 7 .37 (m, 1 H), 7 .16 - 7 .28 (m , 2 H), 7 .06 - 7 .15 (m, 1 H ), 6 .65 - 6 .89 (m, 3 H), 5 .90 (s, 2 H), 2 .59 - 2 .69 (m, 1 H), 1 .54 (s, 3 H), 0.53 - 0.65 (m, 2 H), 0.39 - 0.52 (m, 2 H).

I-423 REFERENCE COMPOSITION

The title compound was prepared following general procedure B, except that 4-(piperidin-4-ylsulfonyl)morpholine (as TFA salt, 1.7 equiv.) was the amine reagent, 4 equivalents of triethylamine were used , and the contents were heated at 105 °C for 12 h. Ethyl acetate was the solvent used for the preparation. The crude material was purified by silica gel chromatography using a gradient of 1 to 5% methanol in dichloromethane over 40 minutes providing the desired compound, reference compound I-423 (32.7 mg, 35% yield) as a solid. white.

1H NMR (500 MHz, CDCh) 8 ppm: 8.48 (d, 1 H), 8.25 (d, 1 H), 7.30 (s, 1 H), 7.20 - 7.25 (m , 1 H), 7.03 - 7.08 (m, 1 H), 6.96 - 7.01 (m, 1 H), 6.83-6.88 (m, 1 H), 6.60 (d , 1 H), 5.98 (s, 2 H), 4.83 (d, 2 H), 3.76 (m, 4 H), 3.41 (m, 4 H), 3.23 -3 .29 (m, 1 H), 3 .06-3 .11 (m, 2 H), 2 .20-2.26 (m, 2 H), 1 .91-2 .03 (m, 2 H) .

Reference compound I-424

The title compound was prepared following general procedure B, except that trans-4-(trifluoromethyl)pyrrolidine-3-carboxylic acid (as the TFA salt, 1.5 equiv.) was the amine reagent, 4 equivalents of triethylamine, and the contents were heated at 105 °C for 3 h. A mixture of dichloromethane:isopropanol (5:1) was used as a solvent for the preparation. Concentration of the dried and vacuum-filtered organic layer provided the desired compound, reference compound I-424 (108.3 mg, 89% yield) as an off-white solid.

1H NMR (500 MHz, CDCla) 8 ppm: 8.50 (d, 1 H), 8.18 (d, 1 H), 7.40 (s, 1 H), 7 .19 -7 .25 (m , 1 H), 6 .98 -7 .07 (m, 3 H), 6.65 (d, 1 H), 5.94 (s, 2 H), 4.26 (m, 1 H), 4 .11 -4 .18 (m, 2 H), 4 .03 -4 .09 (m, 1 H), 3 .48 -3 .55 (m, 1 H), 3 .39 -3 .54 (m , 1 HOUR).

Reference compound I-425

To a suspension of the reference compound I-366 (1 equiv.) in dichloromethane was added cyclopropanecarboxylic acid chloride (1.08 equiv.) and triethylamine (1.1 equiv.). After stirring at room temperature for 24 hours, the reaction remained heterogeneous and incomplete according to LC/MS analysis. An additional 4 equivalents of acid chloride and 1,8-diazabicycloundec-7-ene were added, and the reaction was then heated at 60 °C for 1 h, after which the reaction was tan in color and completely homogeneous. The reaction was diluted in water, acidified by the addition of 1N hydrochloric acid solution, extracted with dichloromethane (3 times), dried (sodium sulfate), filtered and concentrated to give a white solid. Purification was achieved by silica gel using 1 to 8% methanol in dichloromethane for 38 minutes providing the desired compound, reference compound I-425 (40.4 mg, 71% yield) as a white solid.

1H NMR (500 MHz, CDCh) 8 ppm: 8.47 (d, 1 H), 8.23 (d, 1 H), 7.82 (br. s, 1 H), 7.29 (s, 1 H), 7 .17 -7 .23 (m, 1 H), 7.01 - 7.06 (m, 1 H), 6.95 - 6.99 (m, 1 H), 6.81 - 6 .87 (m, 1 H), 6.60 (d, 1 H), 5.96 (s, 2 H), 4.79 - 4.84 (m, 2 H), 3.83 -3.94 (m , 1 H), 3 .08 -3 .15 (m, 2 H), 2 .23 -2 .29 (m, 2 H), 2.01 -2 .06 (m, 1 H), 1.27 (m, 2 H), 1.19 (m, 2 H), 0.99-1.04 (m, 2 H).

Reference compound I-426

To a suspension of the reference compound I-366 (1 equiv.) in dichloromethane was added acetic anhydride (1.25 equiv.) and 1,8-diazabicycloundec-7-ene (1.25 equiv.). After stirring at room temperature for 24 hours, the reaction remained heterogeneous and incomplete as determined by LC/MS. An additional 4 equivalents of acetic anhydride and 1,8-diazabicycloundec-7-ene were added, and the reaction was then heated at 60 °C for 1 h, after which the reaction was tan in color and completely homogeneous. The reaction was diluted in water, acidified by the addition of 1N hydrochloric acid solution, extracted with dichloromethane (3 times), dried (sodium sulfate), filtered and concentrated to give a white solid. Purification was achieved by silica gel chromatography using a gradient of 1 to 8% methanol in dichloromethane over 38 minutes providing the desired compound, reference compound I-426 (24.7 mg, 44% yield) as a white solid. .

1H NMR (500 MHz, CDCh) 8 ppm: 8.47 (d, 1 H), 8.23 (d, 1 H), 7.85 (br. s, 1 H), 7.29 (s, 1 H), 7 .18 -7 .25 (m, 1 H), 7.02 - 7.08 (m, 1 H), 6.96 - 7.02 (m, 1 H), 6.84 - 6 .89 (m, 1 H), 6.61 (d, 1 H), 6.00 (s, 2 H), 4.80 - 4.90 (m, 2 H), 3.85 -3.95 (m , 1 H), 3 .08 -3 .17 (m, 2 H), 2 .23-2 .31 (m, 2 H), 2.17 (s, 3 H), 1 .97 -2 .08 (m, 2H).

Reference compound I-427

The title compound was prepared by general procedure B, except that pyrrolidine-3-sulfonamide (1.35 equiv.) was the amine reagent, 4 equivalents of triethylamine were used, and the contents were heated to 105 °C for 2 hours After cooling, the reaction was poured into a 1:1 mixture of water and ethyl acetate, and the resulting precipitate was filtered and dried under vacuum providing the desired compound, reference compound I-427 (46.7 mg, yield 39%) as a whitish solid.

1H NMR (500 MHz, DMSO-de) 8 (ppm): 9.05 - 9.13 (m, 1 H), 8.24 - 8.31 (m, 1 H), 7.51 - 7.57 (m, 1 H), 7.28 - 7.37 (m, 1 H), 7.18 - 7.27 (m, 2 H), 7.06 - 7.17 (m, 3 H), 6 .76 - 6.87 (m, 1 H), 5.87 - 5.93 (m, 2 H), 4.00 - 4.09 (m, 2 H), 3.84 -3.97 (m, 2 H), 3.75 - 3.84 (m, 1 H), 2.29 - 2.42 (m, 2 H).

I-428 REFERENCE COMPOSITION

The title compound was prepared following general procedure B, except that diethanolamide (1.3 equiv.) was the amine reagent, 2 equiv. of triethylamine was used, and the contents were heated at 100 °C for 12 h. A mixture of dichloromethane:isopropanol (5:1) was used as a solvent for the preparation. The crude material was purified by silica gel chromatography using a gradient of 1 to 8% methanol in dichloromethane over 40 minutes providing the desired compound, reference compound I-428 (24.7 mg, 26% yield) as a solid. white.

1H NMR (500 MHz, CD3OD) 8 (ppm): 8.48 (d, 1 H), 8.17 (d, 1 H), 7.24 (s, 1 H), 7.18 - 7.23 (m, 1 H), 7.01 - 7.06 (m, 1 H), 6.96 - 7.01 (m, 1 H), 6.87-6.92 (m, 1 H), 6 .58 (d, 1 H), 5.95 (s, 2 H), 3.99 (m, 4 H), 3.89 (m, 4 H), 3.30 (br. s, 2 H) . Reference compound I-429

To a solution of the reference compound I-427 (1 equiv.) in dichloromethane was added cyclopropanecarboxylic acid chloride (6 equiv.) followed by 1,8-diazabicycloundec-7-ene (8 equiv.). The reaction was heated for 1 h at 100 °C, after which it was cooled, diluted in water and 1 M hydrochloric acid solution, extracted with dichloromethane (2 times), concentrated and purified directly by gel chromatography. silica with a gradient of 1 to 8% methanol in dichloromethane over 40 minutes providing the desired compound, reference compound I-429 (26.5 mg, 70% yield) as a white solid.

1H NMR (500 MHz, CDCla) 8 ppm: 8.47 (d, 1 H), 8.18 (d, 1 H), 7.31 (s, 1 H), 7 .18 -7 .23 (m , 1 H), 7 .01 -7 .06 (m, 1 H), 6.96 - 7.01 (m, 1 H), 6.83 - 6.88 (m, 1 H), 6.60 (d, 1 H), 5.97 (s, 2 H), 4.40 - 4.47 (m, 2 H), 4.04 - 4.15 (m, 2 H), 3.90 -3 .97 (m, 1 H), 2 .65 -2 .73 (m, 1 H), 2 .45 -2 .53 (m, 1 H), 1.30 (m, 1 H), 1.17 (m, 2 H), 0.99 (m, 2 H).

Reference compound I-430

The title compound was prepared following general procedure B, except that (2R,3R)-2-(hydroxymethyl)pyrrolidin-3-ol (as the TFA salt, 1.7 equiv.) was the amine reagent, 4 equivalents of triethylamine were used, and the contents were heated at 100 °C for 2 h. After cooling, the reaction was diluted with water and methanol, and the resulting precipitate was filtered, washed with water, and dried under vacuum to provide the desired compound, reference compound I-430 (79.3 mg, yield 64 %) as a white solid.

1H NMR (500 MHz, DMSO-d6) 8 (ppm): 9.09 (d, 1 H), 8.24 (d, 1 H), 7.51 (s, 1 H), 7.30 - 7 .36 (m, 1 H), 7.19 - 7.25 (m, 2 H), 7 .08 -7 .13 (m, 1 H), 6 .82 -6 .87 (m, 1 H) , 5.90 (d, 1 H), 5.85 (d, 1 H), 5.23 (d, 1 H), 4.71 (br. s, 1 H), 4 .35 -4 .42 (m, 1 H), 4 .15 -4 .20 (m, 1 H), 3 .77 -3 .87 (m, 2 H), 3 .64 -3 .74 (m, 2 H), 2 .04 -2 .09 (m, 2 H).

Reference compound I-431

The title compound was prepared following general procedure B, except that diisopropanolamine (1 equiv.) was the amine reagent, 4 equivalents of triethylamine were used, and the contents were heated at 100 °C for 24 h. Ethyl acetate was used as a solvent for the preparation. The crude material was purified by silica gel chromatography using a gradient of 1 to 8% methanol in dichloromethane over 40 minutes providing the desired compound, reference compound I-431 (47.1 mg, 42% yield) as a solid. white.

1H NMR (500 MHz, CD3OD) 8 (ppm): 8.72 (d, 1 H), 8.11 (d, 1 H), 7.35 (s, 1 H), 7.23 - 7.28 (m, 1 H), 7.05 - 7.11 (m, 1 H), 7.01 - 7.05 (m, 1 H), 6.83 - 6.88 (m, 2 H, 2 movements isochrons), 5.96 (d, 1 H), 5.92 (d, 1 H), 4.10 - 4.21 (m, 2 H), 3.90 - 3.97 (m, 2 H) , 3.61 - 3.67 (m, 2 H), 1.21 (s, 6 H).

Reference compound I-432

The title compound was prepared in 3 steps:

Step 1: Synthesis of 2-((5-fluoro-2-(1-(2-fluorobenzyl)-5-(isoxazol-3-yl)-1H-pyrazol-3-yl)pyrimidin-4-yl)amino acid )ethanesulfonic

The intermediate product was prepared following general procedure B, except that taurine (1.3 equiv.) was the amine reagent, 2 equiv. of triethylamine was used, and the contents were heated at 100 °C for 8 h. After cooling, the reaction was diluted with 1N HCl solution, filtered, washed with water, and dried in vacuo to provide the desired intermediate, 2-((5-fluoro-2-(1-(2- fluorobenzyl)-5-(isoxazol-3-yl)-1H-pyrazol-3-yl)pyrimidin-4-yl)amino)ethanesulfonic acid (60.2 mg, 64% yield) as a white solid. The intermediate product was used without further purification.

Step 2: Synthesis of 2-((5-fluoro-2-(1-(2-fluorobenzyl)-5-(isoxazol-3-yl)-1H-pyrazol-3-yl)pyrimidin-4-yl) chloride amino)ethanesulfonyl

A suspension of 2-((5-fluoro-2-(1-(2-fluorobenzyl)-5-(isoxazol-3-yl)-1H-pyrazol-3-yl)pyrimidin-4-yl)amino)ethanesulfonic acid (1 equiv.) and thionyl chloride (5 equiv.) in dichloromethane was stirred at room temperature, after which 2 drops of DMF were added. The suspension was heated at 60 °C for 12 hours, after which L C / M analysis indicated that sulfonyl chloride had been formed. The reaction mixture was concentrated by providing 90 mg of a whitish solid.

Stage 3: Synthesis of reference compound I-432

To a suspension of 2-((5-fluoro-2-(1-(2-fluorobenzyl)-5-(isoxazol-3-yl)-1H-pyrazol-3-yl)pyrimidin-4-yl)amino chloride Crude )ethanesulfonyl in tetrahydrofuran was added to an aqueous solution of 50% hydroxylamine (10 equiv.). The reaction became homogeneous instantly, and LC/MS analysis showed it to be complete. The reaction was concentrated to one tert of its volume and purified directly on silica gel using 3 to 10% methanol in dichloromethane for 45 minutes to provide the desired compound, reference compound I-432 (8.2 mg, yield 25%) as a white solid.

1H NMR (500 MHz, CD3OD) 8 (ppm): 8.76 (d, 1 H), 8.13 (d, 1 H), 7.60 (s, 1 H), 7.25 - 7.31 (m, 1 H), 7.07 - 7.13 (m, 1 H), 7.03 - 7.07 (m, 1 H), 6.89 (d, 1 H), 6.83 - 6 .88 (m, 1 H), 6.00 (s, 2 H), 4.07 (t, 2 H), 3.62 (t, 2 H).

Reference compound I-433

The title compound was prepared following general procedure B, except that piperidine-3,5-dione (as the TFA salt, 1.4 equiv.) was the amine reagent, 4 equivalents of triethylamine were used, and the Contents were heated at 100 °C for 1 h. Ethyl acetate was used as a solvent for the preparation. The crude material was purified by silica gel chromatography using a gradient of 3 to 20% methanol in dichloromethane over 40 minutes providing the desired compound, reference compound I-433 (45.6 mg, 44% yield) as a solid. white.

1H NMR (500 MHz, CD3OD) 8 (ppm): 8.77 (s, 1 H), 8.30 (d, 1 H), 7.50 (s, 1 H), 7.26 - 7.32 (m, 1 H), 7.07 - 7.13 (m, 1 H), 7.03 - 7.07 (m, 1 H), 6.94 (d, 1 H), 6.83 - 6 .88 (m, 1 H), 5.98 (s, 2 H), 4.58 (s, 4 H).

Reference compound I-434

The title compound was prepared following general procedure B, except that 4-(hydroxymethyl)piperidin-4-ol (as the TFA salt, 1.4 equiv.) was the amine reagent, 4 equivalents of triethylamine were used , and the contents were heated at 100 °C for 1 h. After cooling, the reaction was diluted with water and 1N HCl solution, and the resulting precipitate was filtered, washed with water, and dried under vacuum to provide the desired compound, reference compound I-434 (88.5 mg , yield 82%) as a white solid.

1H NMR (500 MHz, CD3OD) 8 (ppm): 8.77 (d, 1 H), 8.15 (d, 1 H), 7.43 (s, 1 H), 7.23 - 7.30 (m, 1 H), 7.06 - 7.12 (m, 1 H), 7.00 - 7.05 (m, 1 H), 6.91 (d, 1 H), 6.79 - 6 .84 (m, 1 H), 5.96 (s, 2 H), 4.49 - 4.57 (m, 2 H), 3.49 - 3.57 (m, 2 H), 3.40 (s, 2 H), 1.65 - 1.71 (m, 2 H), 1.77 - 1.84 (m, 2 H).

Reference compound I-435

The title compound was prepared following general procedure B, except that (3R,5S)-5-(hydroxymethyl)pyrrolidin-3-ol (2.7 equiv.) was the amine reagent, 4 equivalents of triethylamine were used , and the contents were heated at 100 °C for 30 min. After cooling, the reaction was diluted with water and methanol, and the resulting precipitate was filtered, washed with water, and dried under vacuum to provide the desired compound, reference compound I-435 (77.3 mg, yield 87 %) as a white solid.

1H NMR (500 MHz, CD3OD) 8 (ppm): 8.75 (d, 1 H), 8.12 (d, 1 H), 7.40 (s, 1 H), 7.24 - 7.28 (m, 1 H), 7.06 - 7.10 (m, 1 H), 7.01 - 7.05 (m, 1 H), 6.90 (d, 1 H), 6.80 - 6 .85 (m, 1 H), 5.95 (s, 2 H), 4.63 - 4.68 (m, 1 H), 4.53 - 4.57 (m, 1 H), 3.81 - 3.94 (m, 3 H), 3 .73 -3 .80 (m, 1 H), 2.10 - 2.25 (m, 2 H).

Reference compound I-437

This compound was prepared in two stages

Stage 1: ketal intermediate

To a suspension of 2-((5-fluoro-2-(1-(2-fluorobenzyl)-5-(isoxazol-3-yl)-1H-pyrazol-3-yl)pyrimidin-4-yl)amino chloride Crude )ethanesulfonyl (synthesis described in step 2 of the procedure to prepare the reference compound I-432 (1 equiv.) in dioxane was added (S)-(2,2-dimethyl-1,3-dioxolan-4-yl )methanamine (2 equiv.). The reaction was stirred for 24 h, after which the reaction mixture was diluted in dichloromethane/isopropanol 5:1 and acidified by the addition of 1 M hydrochloric acid, extracted with dichloromethane/ 5:1 isopropanol (3 x 30 ml), dried (sodium sulfate), filtered and concentrated to give a residue. Purification was achieved by silica gel chromatography using a gradient of 7 to 12% methanol in dichloromethane over. 45 minutes providing the desired compound, reference compound I-43 (31.0 mg, 42% yield) as a white solid.

1 H R M N (500 MHz, C D 3 O D) 8 (pp m): 8, 78 (d, 1 H), 8, 13 (d, 1 H), 7, 52 (s, 1 H), 7, 26 - 7, 32 (m, 1 H), 7, 10 - 7, 15 (m, 1 H), 7, 04 - 7, 08 (m, 1 H), 6, 91 (d, 1 H), 6, 80 - 6.85 (m, 1H), 6.00 (s, 2H), 4.16 - 4.20 (m, 1H), 4.03 - 4.12 (m, 2H), 3.98 - 4.02 (m, 1H), 3.70 - 3.74 (m, 1H), 3.45 - 3.52 (m, 2H), 3.22 (d, 2H) ) , 1 , 34 ( s , 3 H ) , 1 , 27 ( s , 3 H ) .

Stage 2: Composite of reference I - 437

To a solution of the intermediate prepared in step 1 (1 equiv.) in dichloromethane was added a 4 M dioxane solution of hydrogen chloride (20 equiv.). After 1 h, the reaction was concentrated to dryness, diluted with water, extracted with dichloromethane/isopropanol 5:1 (3 times), dried (sodium sulfate), filtered and concentrated to give a residue. Purification was achieved by silica gel chromatography providing the desired compound, reference compound I-437 (10.3 mg, 38.2% yield) as an off-white solid.

1H NMR (500 MHz, CD3OD) 8 (ppm): 8.77 (d, 1 H), 8.12 (d, 1 H), 7.53 (s, 1 H), 7.26 - 7.32 (m, 1 H), 7.08 - 7.13 (m, 1 H), 7.03 - 7.08 (m, 1 H), 6.93 (d, 1 H), 6.81 - 6 .86 (m, 1 H), 6.00 (s, 2 H), 4.03 - 4.13 (m, 2 H), 3.70 - 3.75 (m, 1 H), 3.47 - 3.54 (m, 3 H), 3.24 - 3.29 (m, 2 H), 3.08 - 3.13 (m, 1 H).

Reference compound I-438

The title compound was prepared following general procedure B, except that 4-hydroxypiperidine-4-carboxamide (1.4 equiv.) was the amine reagent, 4 equivalents of triethylamine were used, and the contents were heated to 100° C for 30 min. Ethyl acetate was used as a solvent for the preparation, and purification was achieved by precipitation of the product by trituration of an acetonitrile solution of the crude product with water. The resulting precipitate was filtered and dried under vacuum to provide the desired compound, reference compound I-438 (26.3 mg, 26% yield) as a white solid.

1H NMR (500 MHz, DMSO-de) 8 (ppm): 9.08 (d, 1 H), 8.29 (d, 1 H), 7.55 (s, 1 H), 7,30-7 .35 (m, 2H, 2 isochronous displacements), 7.26 (d, 2 H), 7.19 - 7.25 (m, 1 H), 7.16 (br. s, 1 H), 7, 08 - 7.13 (m, 1 H), 6.79 - 6.84 (m, 1 H), 5.90 (s, 2 H), 4.39 (m, 2 H), 3 .34 - 3 .39 (m, 2 H), 1.95-2.01 (m, 2 H), 1.58 (m, 2 H).

Reference compound I-439

To a suspension of 2-((5-fluoro-2-(1-(2-fluorobenzyl)-5-(isoxazol-3-yl)-1H-pyrazol-3-yl)pyrimidin-4-yl)amino chloride )ethanesulfonyl (synthesis described in step 2 of the procedure for the reference compound I-432, 1 equiv.) in dioxane ethanolamine (2.2 equiv.) was added. The reaction was stirred for 24 h at room temperature, after which the reaction mixture was diluted in 5:1 dichloromethane/isopropanol and acidified by the addition of 1 M hydrochloric acid, extracted with 5:1 dichloromethane/isopropanol (3 times), dried (sodium sulfate), filtered and concentrated to give a residue. Purification was achieved by silica gel chromatography using a gradient of 3 to 8% methanol in dichloromethane over 45 minutes providing the desired compound, reference compound I-439 (31.3 mg, 65% yield) as a white solid. .

1H NMR (500 MHz, DMSO-de) 8 (ppm): 9.09 (d, 1 H), 8.24 (d, 1 H), 7.78 - 7.83 (m, 1 H), 7 .54 (s, 1 H), 7.30 - 7.35 (m, 1 H), 7.21 -7 .25 (m, 2 H), 7.17 (d, 1 H), 7 .08 -7 .13 (m, 1 H), 6.81 -6 .86 (m, 1 H), 5.90 (s, 2 H), 4.80 (t, 1 H), 3.84 (q , 2 H), 3.44 (q, 2 H), 3.39 (t, 2 H), 3.03 (q, 2 H).

Reference compound I-440

The title compound was synthesized in 2 steps:

Stage 1: Synthesis of 1-(benzyloxy)-N-(1-(5-fluoro-2-(1-(2-fluorobenzyl)-5-

-(isoxazol-3-yl)-1H-pyrazol-3-yl)pyrimidin-4-yl)azetidin-3-yl)cyclopropanecarboxamide

The intermediate product was prepared following general procedure B, except that N-(acetidin-3-yl)-1-(benzyloxy)cyclopropanecarboxamide (as the TFA salt, 1.2 equiv.) was the amine reagent, used 4 equivalents of triethylamine, and the contents were heated at 100 °C for 2 h. After cooling, the reaction mixture was diluted with water, and the resulting precipitate was filtered, washed with water, and dried under vacuum to provide the desired intermediate, 1-(benzyloxy)-N-(1-(5- fluoro-2-(1-(2-fluorobenzyl)-5-(isoxazol-3-yl)-1H-pyrazol-3-yl)pyrimidin-4-yl)azetidin-3-yl)cyclopropanecarboxamide (104.3 mg, yield 86%) as a white solid.

Stage 2: Synthesis of the reference compound I-440

To a suspension of 1-(benzyloxy)-N-(1-(5-fluoro-2-(1-(2-fluorobenzyl)-5-(isoxazol-3-yl)-1H-pyrazol-3-yl)pyrimidin -4-yl)azetidin-3-yl)cyclopropanecarboxamide in ethanol was added to 10% palladium on carbon (0.1 equiv). The reaction mixture was evacuated to dryness and a hydrogen balloon was applied. For solubility purposes, ethyl acetate was also added. After 24 h, a mixture of starting material, product and other by-products was observed. The reaction mixture was filtered through celite, then purified by silica gel chromatography using a gradient of 1 to 8% dichloromethane in methanol over 60 minutes providing the desired compound, reference compound I-440 (3, 8 mg, yield 5%). Several subsequent fractions containing the desired product were contaminated with a nearby byproduct and were discarded.12

1H NMR (400 MHz, CDCla) 8 (ppm): 8.44 (d, 1 H), 8.09 (d, 1 H), 7.92 (br. s, 1 H), 7.20 - 7 .25 (m, 1 H), 6.93 - 7.03 (m, 2 H, 2 isochronous transitions), 6.83 - 6.88 (m, 2 H), 6.56 (d, 1 H) , 5.94 (s, 2 H), 4.81 - 4.91 (m, 1 H), 4.55 - 4.63 (m, 1 H), 4 .05 -4 .12 (m, 2 H), 2.31 -2 .36 (m, 1 H), 1 .33 - 1.37 (m, 2 H), 1 .04 - 1.09 (m, 2 H).

REFERENCE COMPOSITION I - 441

The title compound was prepared following general procedure B, except that 3-(aminomethyl)-1H-1,2,4-triazol-5(4H)-one (1.3 equiv.) was the amine reagent, 4 equivalents of triethylamine were used and the contents were heated at 100 °C for 12 h. After cooling, the reaction was filtered and purified by silica gel chromatography using a gradient of 3 to 10% methanol in dichloromethane over 40 minutes to provide the desired compound, reference compound I-441 (12.9 mg, yield 13%) as a white solid.

1H NMR (500 MHz, CD3OD) 8 (ppm): 8.78 (d, 1 H), 8.17 (m, 1 H), 7.42 (s, 1 H), 7.25 - 7.31 (m, 1 H), 7.07 - 7.12 (m, 1 H), 7.02 - 7.07 (m, 1 H), 6.88 - 6.93 (m, 2 H, 2 transitions isochronas), 6.02 (s, 2 H), 4.61 (s, 2 H).

Reference compound I-442

To a suspension of 1,2,4-triazolin-3-one (0.5 equiv.), potassium carbonate (1.5 equiv.), and copper (I) iodide (0.05 equiv.) in W To/-dimethylformamide, intermediate 1 (1 equiv.) was added followed by trans-1,2-bis-(methylamino)cyclohexane (0.1 equiv.). The reaction was heated at 100 °C for 24 h, after which the reaction was diluted in water, extracted with ethyl acetate, dried (sodium sulfate), filtered, and concentrated. Two purification attempts were carried out by silica gel chromatography (5-7% methanol in dichloromethane), giving rise to a peak corresponding to three compounds. Further purification by reverse phase HPLC (5 to 75% acetonitrile in water with addition of 0.1% trifluoacetic acid for 20 minutes) provided the desired compound, reference compound I-442 (0.9 mg, yield 0 .8%) as a white solid. 1H NMR (500 MHz, CD3OD) 8 (ppm): 8.96 (s, 1 H), 8.81 (d, 1 H), 8.12 (s, 1 H), 7.66 (s, 1 H), 7.25 - 7.31 (m, 1 H), 7.09 - 7.15 (m, 1 H), 7.02 - 7.07 (m, 1 H), 6.89 - 6 .94 (m, 1 H), 5.98 (s, 2 H).

Reference compound I-443

The title compound was prepared following general procedure B, except that 3-amino-2-hydroxypropanamide (2 equiv.) was the amine reagent, 4 equivalents of triethylamine were used, and the contents were heated to 110 °C for 6 hrs. A mixture of dichloromethane:isopropanol (5:1) was used as a solvent for the preparation. The crude material was purified by silica gel chromatography using a gradient of 3 to 10% methanol in dichloromethane over 45 minutes providing the desired compound, reference compound I-443 (17.3 mg, 14% yield) as a solid. white.

1H NMR (500 MHz, CD3OD) 8 (ppm): 8.78 (d, 1 H), 8.11 (d, 1 H), 7.52 (s, 1 H), 7 .27-7 .31 (m, 1 H), 7 .09 -7 .13 (m, 1 H), 7.03 - 7.08 (m, 1 H), 6.95 (d, 1 H), 6.83 - 6 .88 (m, 1 H), 5.99 (s, 2 H), 4.37 (dd, 1 H), 4.05 (dd, 1 H), 3.81 (dd, 1 H).

Reference compound I-444

To a solution of intermediate 2 (intermediate described in patent application publication WO2012/3405 A1) (1 equiv.) in dichloromethane was added 2,2,2-trifluoroethanesulfonyl chloride (1.08 equiv.) followed by 1,8-diazabicycloundec-7-ene (1.2 equiv.). The reaction was stirred for 16 h at 23 °C, after which the reaction mixture was diluted in water and 1N hydrochloric acid solution, extracted with ethyl acetate (3 times), washed with 1N hydrochloric acid solution (2 times), dried (sodium sulfate), filtered and concentrated in vacuo. Purification of the crude material by silica gel chromatography using isocratic gradient of 5% methanol in dichloromethane provided the desired compound, reference compound I-444 (28.9 mg, 27% yield) as an off-white solid.12

1H NMR (500 MHz, CD3OD) 8 (ppm): 8.82 (d, 1 H), 8.24 (br. s, 1 H), 7.57 (s, 1 H), 7.26 - 7 .32 (m, 1 H), 7.08 - 7.13 (m, 1 H), 7.03 - 7.08 (m, 1 H), 6.93 - 6.99 (m, 2 H, 2 overlapping movements), 6.79 - 6.85 (br. s, 1 H), 6.01 (s, 2 H), 4.49 - 4.58 (m, 2 H).

Reference compound I-445

To a suspension of sodium hydride (1.2 equiv.) in anhydrous tetrahydrofuran at 23 °C, intermediate-2 (1 equiv.) was added. The reaction was left stirring for 30 min at 23 °C. 3,3,3-Trifluoropropane-1-sulfonyl chloride (1 equiv.) dissolved in tetrahydrofuran was added to the reaction mixture, which was stirred for 18 h. The reaction mixture was diluted in water and 1N hydrochloric acid solution, extracted with ethyl acetate (3 times), washed with 1N hydrochloric acid solution (2 times), dried (sodium sulfate), filtered and concentrated in vacuo. Purification of the crude material by reverse phase HPLC provided the desired compound, reference compound I-445 (17.1 mg, 20% yield) as an off-white solid.

1H NMR (500 MHz, CDCh) 8 ppm: 8.51 (d, 1 H), 8.25 (br. s, 1 H), 7.20 -7.29 (m, 2 H), 6.98 - 7.08 (m, 3 H), 6.59 (d, 1 H), 5.80 - 5.99 (m, 2 H), 3.64 (br. s, 2 H), 2.69 - 2.88 (m, 2 H).

Reference compound I-446

To a suspension of sodium hydride (1.2 equiv.) in anhydrous tetrahydrofuran at 23 °C was added the intermediate product -21 equiv.). The reaction was left stirring for 30 min at 23 °C. 2-Methoxyethanesulfonyl chloride (1 equiv.) dissolved in tetrahydrofuran was dissolved into the reaction mixture, which was stirred for 18 h. The reaction mixture was diluted in water and 1N hydrochloric acid solution, extracted with ethyl acetate (3 times), washed with 1N hydrochloric acid solution (2 times), dried (sodium sulfate), filtered and he concentrated. Purification of the crude material by reverse phase HPLC provided the desired compound, reference compound I-446 (9.1 mg, 13.5% yield) as an off-white solid.

1H NMR (500 MHz, CDCla) 8 ppm: 8.50 (d, 1 H), 8.37 (br. s, 1 H), 7.19-7.27 (m, 2 H), 6.92 - 7.07 (m, 3 H), 6.61 (d, 1 H), 5.94 (s, 2 H), 3.85 - 3.92 (m, 2 H), 3.78 (br .s, 2H), 3.28 (s, 3H).

Reference compound I-447

Oxalyl chloride (4 equiv.) was added to a solution of triethylamine (4 equiv.) and the reference compound I-214 (1 equiv.) in DCM was kept at 0 °C. The reaction was warmed and stirred at room temperature for 2 h. The reaction was then quenched by the addition of water, extracted with dichloromethane, and the organic extracts were concentrated in vacuo to provide the desired compound, reference compound I-447 (3.7 mg, 69% yield) as a solid. .

1H-NMR (500 MHz, DMSO-d6) 8 ppm 9.11 (d, 1 H), 8.90 (d, 1 H), 7.65 (s, 1 H), 7.30-7.37 (m, 1 H), 7.27 (d, 1 H), 7.19 - 7.26 (m, 1 H), 7.11 (t, 1 H), 6.87 (t, 1 H) , 5.93 (s, 2 H), 4 .00-4.07 (m, 2 H), 2.57 (t, 2 H), 2.13-2.21 (m, 2 H).

Reference compound I-448

The title compound was prepared following general procedure B, except that 5-(aminomethyl)thiophen-2-carboxylic acid (4 equiv.) was the amine reagent, 9 equivalents of triethylamine were used, and the contents were heated to 110 °C as a solution in dioxane/water (4.5:1) for 2 d. The contents were diluted with ethyl acetate and 1N HCl solution, and the resulting precipitate was filtered. The solids were collected and dried under vacuum to provide the desired compound, reference compound I-448 (12 mg, 27% yield) as a solid. 1H R<m>N (500 MHz, DMSO-cfe) 8 ppm 12.9 (br. s., 1 H), 9.11 (d, 1 H), 8.92 - 9.08 (m, 1 H), 8.35 (t, 1 H), 7.61 - 7.67 (m, 1 H), 7.56 (d, 1 H), 7.30 - 7.38 (m, 1 H) , 7.19 - 7.26 (m, 3 H), 7.12 (t, 1 H), 6.92 (t, 1 H), 5.92 (s, 2 H), 4.88 - 4 .97 (m, 2H).

Reference compound I-449

The title compound was prepared following general procedure B, except that 5-aminopentanoic acid (4 equiv.) was the amine reagent, 9 equivalents of triethylamine were used, and the contents were heated to 110 °C as a solution in dioxane/water (4.5:1) for 2 d. The contents were diluted with ethyl acetate and 1N HCl solution, and the resulting precipitate was filtered. The solids were collected and dried under vacuum to provide the desired compound, reference compound I-449 (34 mg, 84% yield) as a solid.

1H NMR (500 MHz, DMSO-d6) 8 ppm 12.01 (s, 1 H), 9.12 (s, 1 H), 8.32 - 8.62 (m, 1 H), 8.27 ( br. s., 1 H), 7.61 (br. s., 1 H), 7.29 - 7.40 (m, 1 H), 7.18 - 7.27 (m, 2 H), 7.11 (t, 1 H), 6.88 (t, 1 H), 5.93 (br. s., 2 H), 2.29 (t, 3 H), 1.49 - 1.72 (m, 5 H).

Reference compound I-450

A suspension of 5-fluoro-2-(1 -(2-fluorobenzyl)-5-(isoxazol-3-yl)-1H-pyrazol-3-yl)—pyrimidin-4-ol (generated by general procedure A, using 1-(isoxazol-3-yl)ethanone in step 1 and 2-fluorobenzylhydrazine in step 2) (1 equiv.) and sodium methoxide in methanol (0.5 M solution, 4 equiv.) was heated in a Microwave container at 130°C for 4 h. The reaction was quenched with 1N HCl solution at pH 2, and the resulting residue was filtered. The solids were washed with methanol and dried under vacuum to provide the desired compound, reference compound I-450 (1.45 g, 68%) as a white solid.

1H NMR (500 MHz, CD3OD) 8 ppm 8.04 (d, 1 H), 7.71 (s, 1 H), 7.23 - 7.36 (m, 1 H), 7.00 - 7, 18 (m, 2 H), 6.90 (t, 1 H), 5.94 (s, 2 H), 2.56 (s, 3 H)

Reference compound I-451

The composition of the title was prepared in 2 stages:

Step 1: Synthesis of 1-(3-(4-chloro-5-fluoropyrimidin-2-yl)-1-(2-fluorobenzyl)-1H-pyrazol-5-yl)ethanone

Reference compound I-450 was loaded with phosphoryl trichloride (60 equiv.) and the resulting mixture was stirred at 45 °C until the reaction was considered complete as determined by L<c>/MS. The reaction was then carefully poured onto ice, extracted with 4:1 dichloromethane/isopropanol and the layers separated. The organic parts were combined, dried over sodium sulfate, filtered, and concentrated in vacuo. The material was transferred to the next step without further purification.

Stage 2: Synthesis of reference compound I-451

The title compound was prepared following general procedure B, except that 1-((methylamino)methyl)cyclopropanecarboxylic acid was the amine reagent, 1-(3-(4-chloro-5-fluoropyrimidin-2-yl) was used )-1-(2-fluorobenzyl)-1H-pyrazol-5-yl)ethanone instead of intermediate 1, and the content was heated at 100 °C for 36 h as a solution in dioxane. The crude material was purified by reverse phase HPLC providing the desired compound, reference compound I-451 (50 mg, 69% yield) as a tan solid.

1H NMR (500 MHz, DMSO-da) 8 ppm 12.53 (br. s, 1 H), 8.19 (d, 1 H), 7.65 (s, 1 H), 7.33 (d, 1 H), 7.17 - 7.26 (m, 1 H), 7.11 (t, 1 H), 6.86 (t, 1 H), 5.81 (s, 2 H), 4, 00 (s, 2 H), 3.24 (d, 3 H), 2.57 (s, 3 H), 1.03 (d, 2 H), 0.74-0.91 (m, 2 H ).

Reference compound I-452 and reference compound I-453

The title compound was prepared following general procedure B, except that 4-(trifluoromethyl)piperidine-2-carboxylic acid was the amine reagent, Hunig's base (8 equiv.) was used instead of triethylamine, and the content was heated at 120 °C for 18 h as a solution in THF/water (1:1). The solvent was removed in vacuo and the resulting residue was purified by reverse phase HPLC, then with a gradient of 0-10% methanol/dichloromethane providing the desired compounds, reference compound I-452 (14 mg, 15% yield). ) as a solid and the reference compound I-453 (18.4 mg, yield 21%) as a solid. The assignment of trans versus cis was done arbitrarily at the time of synthesis; each diastereomer is a mixture of racemates.

1H NMR of Reference Compound I-452 (500 MHz, CD3OD) 88.72 - 8.79 (m, 1 H), 8.20 - 8.25 (m, 1 H), 7.41 - 7.47 (m, 1 H), 7.22 - 7.32 (m, 1 H), 6.99 - 7.14 (m, 2 H), 6.87 - 6.92 (m, 1 H), 6 .76 - 6.84 (m, 1 H), 5.93 - 5.99 (m, 2 H), 5.59 - 5.76 (m, 1 H), 4.64 - 4.80 (m , 1 H), 3.38 - 3.46 (m, 1 H), 2.55 - 2.63 (m, 1 H), 2.40 - 2.54 (m, 1 H), 2.00 - 2.08 (m, 1 H), 1.84 - 1.96 (m, 1 H), 1.64 - 1.75 (m, 1 H).

1H NMR of Reference Compound I-453 (500 MHz, CD3OD) 88.73 - 8.79 (m, 1 H), 8.23 - 8.33 (m, 1 H), 7.43 (s, 1 H), 7.24 - 7.31 (m, 1 H), 7.07 - 7.13 (m, 1 H), 7.00 - 7.06 (m, 1 H), 6.82 - 6 .88 (m, 2 H), 5.96 (s, 2 H), 4.52 - 4.60 (m, 1 H), 4 .05-4.14 (m, 1 H), 3.75 -3.85 (m, 1 H), 2 .64-2.78 (m, 1 H), 2.41 -2.49 (m, 1 H), 2.12 (dd, 2 H), 1 .77- 1.87 (m, 1 H).

Reference compound I-454

The title compound was prepared by general procedure B, except that 3-(trifluoromethyl)piperidine-2-carboxylic acid (5 equiv.) was the amine reagent, and the contents were heated at 90 °C for 18 h as a solution in THF/water (3:1). The solvent was removed in vacuo and the resulting residue was purified by reverse phase HPLC, then with a gradient of 0-15% methanol/dichloromethane providing the desired compound, reference compound I-454 (1.6 mg, yield 4%) as a solid. 1H<r>M<n>(500 MHz, CD3OD) 8 ppm 8.76 - 8.80 (m, 1 H), 8.28 - 8.33 (m, 1 H), 7.49 (s, 1 H), 7.25 - 7.32 (m, 1 H), 7.02 - 7.13 (m, 2 H), 6.84 - 6.90 (m, 2 H), 5.94 - 6.00 (m, 2 H), 5.37 - 5.41 (m, 1 H), 4.35 - 4.43 (m, 1 H), 3.80 - 3.89 (m, 1 H) ), 2.89 - 3.00 (m, 1 H), 1.93 - 2.08 (m, 3 H), 1.77 - 1.90 (m, 1 H).

Reference compound I-455

The title compound was prepared following general procedure B, except that 3-ethylpiperidine-2-carboxylic acid (1 equiv.) was the amine reagent, and the contents were heated at 90 °C for 18 h as a solution in THF/water (5:1). The solvent was removed in vacuo and the resulting residue was purified by reverse phase HPLC to provide the desired compound, reference compound I-455 (5 mg, 15% yield) as a solid.

1H NMR (500 MHz, CD3OD) 8 ppm 8.81 - 8.86 (m, 1 H), 8.33 - 8.40 (m, 1 H), 7.53 - 7.59 (m, 1 H) ), 7.27 - 7.35 (m, 1H), 7.03 - 7.16 (m, 2H), 6.91 - 7.00 (m, 2H), 5.99 - 6.05 (m, 2 H), 5.39 - 5.47 (m, 1 H), 4.55 - 4.68 (m, 1 H), 3.78 - 3.89 (m, 1 H), 1 .75-2.06 (m, 5 H), 1.46- 1.60 (m, 2H), 1.07-1.13 (m, 3 H).

Reference compound I-362

The title compound was prepared following general procedure B, except that 1-(3-(4-chloro-5-fluoropyrimidin-2-yl)-1-(2-fluorobenzyl)-1H-pyrazole-5- yl)ethanone (described in step 1 for the synthesis of reference compound I-451 instead of intermediate 1, 2-(aminomethyl)-1,1,1,3,3,3-hexafluoropropan-2-ol was the amine reagent and the content was heated at 100 °C for 36 h as a solution in dioxane. The resulting crude material was purified by silica gel chromatography with a gradient of 0-10% methanol/dichloromethane followed by.

1H-NMR (500 MHz, DMSO-cfe) 8 ppm 8.54 (s, 1 H), 8.36 (d, 1 H), 8.07 (br. s., 1 H), 7.58 - 7.69 (m, 1 H), 7.33 (q, 1 H), 7.16 - 7.26 (m, 1 H), 7.10 (t, 1 H), 6.92 - 7, 02 (m, 1 H), 5.81 (s, 2 H), 4.12 (d, 2 H), 2.56 (s, 3 H).

Reference compound I-462

The title compound was prepared following general procedure B, except that 1-(3-(4-chloro-5-fluoropyrimidin-2-yl)-1-(2-fluorobenzyl)-1H-pyrazole-5- yl)ethanone (described in step 1 for the synthesis of reference compound I-451 instead of intermediate 1, 2-aminopropane-1,3-diol was the amine reagent and the content was heated to 100 °C for 36 h as a solution in dioxane. The resulting crude material was purified by silica gel chromatography with a gradient of 0-10% methanol/dichloromethane followed by further purification by reverse phase HPLC providing the desired compound, the reference compound. I-462 (10 mg, 26% yield) as an off-white solid.

1H-NMR (500 MHz, DMSO-d6) 8 ppm 8.18 (d, 1 H), 7.65 (d, 1 H), 7.33 (q, 1 H), 7.21 (dd, 2 H), 7.10 (t, 1 H), 6.79 (t, 1 H), 5.82 (s, 2 H), 4.72 (t, 2 H), 4.33 (d, 1 H), 3.52 - 3.64 (m, 4 H), 2.57 (s, 3 H).

Reference compound I-463

The title compound was prepared following general procedure B, except that 1-(3-(4-chloro-5-fluoropyrimidin-2-yl)-1-(2-fluorobenzyl)-1H-pyrazole-5- yl)ethanone (described in step 1 for the synthesis of reference compound I-451 instead of intermediate 1, (2R,3S)-3-methylpiperidine-2-carboxylic acid was the amine reagent and the content was heated at 100 °C for 36 h as a solution in dioxane. The crude material was purified by silica gel chromatography with a gradient of 0-10% methanol/dichloromethane followed by further purification by reverse phase HPLC to provide the desired compound, the reference compound I-463 (6.5 mg, 15% yield) as an off-white solid.

1H-NMR (500 MHz, DMSO-d6) 8 ppm 8.38 (d, 1 H), 7.69 (s, 1 H), 7.33 (q, 1 H), 7.17-7.27 (m, 1 H), 7.11 (t, 1 H), 6.77 - 6.89 (m, 1 H), 5.82 (s, 2 H), 4.85 (d, 1 H) , 4.26 - 4.28 (m, 1 H), 3.48 - 3.57 (m, 1 H), 2.56 (s, 3 H), 1.98 - 2.12 (m, 1 H), 1.83 (d, 1 H), 1.58-1.71 (m, 2 H), 1.34- 1.47 (m, 1 H), 1.11 (d, 3 H) , COOH exchanged protons.

Reference compound I-519

2-((ted-butoxycarbonyl)amino)acetic acid (1 equiv.), HATU (1.1 equiv.), and Hunig's base (1.3 equiv.) were stirred as a solution in DMF at 23 °C for 2 hours Intermediate 2 (1 equiv.) was then added in one portion and the contents were microwaved at 100 °C for 30 min. The reaction was quenched by the addition of water, extracted with ethyl acetate, and the organic extracts were washed with water and brine. The mixture was dried, filtered, concentrated in vacuo and purified by silica gel chromatography to provide the desired compound, reference compound I-519 (60 mg, 53% yield) as a yellow solid.

1H-NMR (500 MHz, DMSO-d6) 8 ppm 11.03 - 11.19 (m, 1 H), 9.11 (d, 1 H), 8.73 (d, 1 H), 7 .93 -8.04 (m, 1 H), 7.65 (s, 1 H), 7.33 (q, 1 H), 7.20 - 7.29 (m, 2 H), 7.06 - 7 .16 (m, 2 H), 6.89 (t, 1 H), 5.92 (s, 2 H), 3.83 (d, 2 H), 1.36 - 1.42 (m, 9 H).

Reference compound I-520

To a solution of reference compound I-519 in dichloromethane at 0 °C was added trifluoacetic acid of equal volume and the contents were allowed to warm to 23 °C over a period of 12 h. The contents were dried in vacuo and the resulting residue was taken up in a saturated NaHCO3 solution and extracted with an isopropanol/dichloromethane mixture. The contents were dried, filtered and concentrated in vacuo to provide the desired compound, reference compound I-520 (37 mg, 79% yield) as a brown solid.

1H-NMR (400 MHz, DMSO-d6) 8 ppm 9.10 (d, 1 H), 8.74 (d, 1 H), 8.04 (d, 1 H), 7.68 (s, 1 H), 7.34 (q, 1 H), 7.29 (d, 1 H), 7 .18-7.26 (m, 1 H), 7.11 (t, 1 H), 6.88 (t, 1 H), 5.93 (s, 2 H), 3.36 (s, 2 H), NH protons were exchanged.

Reference compound I-535

To a solution of reference compound I-520 (1 equiv.) and triethylamine (2 equiv.) in dichloromethane maintained at 0 °C was added acetyl chloride (1.2 equiv.), and the resulting mixture was allowed to warm until 23 °C for a period of 18 hours. The solvents were removed in vacuo and the resulting residue was triturated with an ethyl acetate/hexanes mixture. The content was filtered, and the resulting solid was dried under vacuum to provide the desired compound, reference compound I-535 (6 mg, 53% yield) as a white solid.1

1H-NMR (500 MHz, DMSO-d6) 8 ppm 11.18 (s, 1 H), 9.11 (d, 1 H), 8.73 (d, 1 H), 8.22 (t, 1 H), 7.98 (d, 1 H), 7.66 (s, 1 H), 7.30-7.39 (m, 1 H), 7.27 (d, 1 H), 7.20 - 7.26 (m, 1 H), 7.12 (td, 1 H), 6.89 (t, 1 H), 5.93 (s, 2 H), 3.97 (d, 2 H) , 1.83 - 1.93 (m, 3 H).

REFERENCE COMPOSITION I - 543

To a solution of reference compound I-520 (1 equiv.) and triethylamine (6 equiv.) in dichloromethane maintained at 0 °C was added methanesulfonyl chloride (3.3 equiv.), and the resulting mixture was allowed to warm to 23 °C for a period of 3 h. The reaction was quenched by addition of water, and the mixture was extracted with dichloromethane. The contents were dried, concentrated in vacuo and purified by silica gel chromatography to provide the desired compound, reference compound I-543 (6.2 mg, 47% yield) as a white solid.

1H-NMR (500 MHz, DMSO-da) 8 ppm 11.17 (s, 1 H), 9.11 (d, 1 H), 8.76 (d, 1 H), 8.00 (d, 1 H), 7.66 (s, 1 H), 7.54 (t, 1 H), 7.30 - 7.39 (m, 1 H), 7.27 (d, 1 H), 7.21 - 7.26 (m, 1 H), 7.12 (t, 1 H), 6.89 (t, 1 H), 5.93 (s, 2 H), 3.99 (d, 2 H) , 2.98 (s, 3 H).

Reference compound I-584

To a solution of the reference compound I-520 (1 equiv.) in dichloromethane was added isocyanatotrimethylsilane (1.1 equiv.), and the resulting suspension was heated at 40 °C for 18 h. After completion of the reaction, the solvent was removed in vacuo and the resulting residue was purified by reverse phase HPLC to provide the desired compound, reference compound I 584 (16 mg, 55% yield) as a white solid.

1H-NMR (500 MHz, DMSO-d6) 8 ppm 11.02 (s, 1 H), 9.10 (d, 1 H), 8.73 (d, 1 H), 7.99 (d, 1 H), 7.59 - 7.68 (m, 1 H), 7.30 - 7.37 (m, 1 H), 7.19 - 7.27 (m, 2 H), 7.12 (t , 1 H), 6.89 (t, 1 H), 6.25 (t, 1 H), 5.93 (s, 2 H), 5.72 (s, 2 H), 3.90 (d , 2H).

Reference compound I-585

To a solution of the reference compound I-520 (1 equiv.) in dichloromethane was added isopropyl isocyanate (1.1 equiv.), and the resulting mixture was heated at 40 °C for 18 h. After completion of the reaction, the solvent was removed in vacuo and the resulting residue was purified by reverse phase HPLC to provide the desired compound, reference compound I 585 (19 mg, 59% yield) as a white solid.

1H-NMR (500 MHz, DMSO-d6) 8 ppm 11.04 (s, 1 H), 9.10 (d, 1 H), 8.72 (d, 1 H), 7.98 (d, 1 H), 7.64 (s, 1 H), 7.30-7.38 (m, 1 H), 7.19 - 7.28 (m, 2 H), 7.12 (td, 1 H) , 6.85 - 6.95 (m, 1 H), 6.09 (d, 1 H), 6.01 (t, 1 H), 5.92 (s, 2 H), 3.92 (d , 2 H), 3.58-3.71 (m, 1 H), 1.03 (d, 6 H).

Reference compound I-586

To a solution of the reference compound I-520 (1 equiv.) and triethylamine (2 equiv.) in dichloromethane was added dimethylsulfamoyl chloride (1.5 equiv.), and the mixture was heated at 40 °C for 18 h. After completion of the reaction, the solvents were removed in vacuo and the resulting residue was purified by reverse phase HPLC to provide the desired compound, reference compound I-586 (9 mg, 28% yield) as a white solid.

1H-NMR (500 MHz, DMSO-d6) 8 ppm 11.12 (s, 1 H), 9.10 (d, 1 H), 8.75 (d, 1 H), 8.00 (d, 1 H), 7.64 (s, 1 H), 7.58 (t, 1 H), 7.30 - 7.38 (m, 1 H), 7.19 - 7.27 (m, 2 H) , 7.12 (td, 1 H), 6.90 (t, 1 H), 5.92 (s, 2 H), 3.92 (d, 2 H), 2.67 (s, 6 H) .

Reference compound I-633

The title compound was prepared following general procedure B, except that (3-(trifluoromethyl)-5,6,7,8-tetrahydro-[1,2,4]triazolo[4,3-a]pyrazin-6 -yl)methanol was the amine reagent, and the content was heated at 100 °C for 48 h as a solution in THF/dioxane/water (1:10:1). The crude material was purified by silica gel chromatography with a gradient of 0-10% methanol/dichloromethane followed by reverse phase HPLC providing the desired compound, reference compound I-633 (6 mg, 36% yield) as a whitish solid.

1H-NMR (500 MHz, DMSO-d6) 8 ppm 9.11 (d, 1 H), 8.46 (d, 1 H), 7.67 (s, 1 H), 7.30-7.38 (m, 1 H), 7.28 (d, 1 H), 7.19 - 7.25 (m, 1 H), 7.11 (t, 1 H), 6.85 (t, 1 H) , 5.88- 5.98 (m, 2 H), 5.61 (d, 1 H), 5.20 (t, 1 H), 5.13 (br. s., 1 H), 4, 82 (d, 1 H), 4.38 - 4.51 (m, 2 H), 3.64 (dt, 1 H), 3.50 - 3.59 (m, 1 H).

Reference compound I-466

The title compound was prepared following general procedure B, except that 2,2,2-trifluoroethanamine (as the HCl salt) was the amine reagent, and the contents were heated at 90–100 °C for 42 h. The crude material was purified by preparative HPLC using a gradient of 25-80% acetonitrile/water (with 0.1% TFA) providing the desired compound, reference compound I-466 (35 mg, 60% yield) as a solid. white.

1H-NMR (500 MHz, CD3OD) 8 ppm 8.76 (d, 1 H), 8.20 (d, 1 H), 7.46 (s, 1 H), 7.28 (ap. q, 1 H), 7.10 (m, 1 H), 7.03 (ap. t, 1 H), 6.92 (d, 1 H), 6.80 (ap. t, 1 H), 5.97 (s, 2 H), 4.44 (q, 2 H).

REFERENCE COMPOSITION I - 487

The title compound was prepared following general procedure B, except that 2-(methylsulfonyl)ethanamine (as the HCl salt) was the amine reagent, and the contents were heated at 100 °C for 17 h. The contents were cooled to room temperature, diluted with water and acidified to a pH of 3 with a 1N HCl solution. The resulting precipitate was filtered and dried under vacuum to provide the desired compound, reference compound I-487 (56 mg, 91% yield) as a white solid.

1H-NMR (500 MHz, DMSO-da) 8 ppm 9.10 (d, 1 H), 8.24 (d, 1 H), 7.93 (br. t, 1 H), 7.54 (s , 1 H), 7.33 (ap. q, 1 H), 7.22 (m, 1 H), 7.18 (d, 1 H), 7.11 (ap. t, 1 H), 6 .88 (ap. t, 1 H), 5.89 (s, 2 H), 3.87 (dt, 2 H), 3.46 (t, 2 H), 3.06 (s, 3 H) .

Reference compound I-502

The title compound was prepared following general procedure B, except that (1-aminocyclopropyl)methanol (as the HCl salt) was the amine reagent and the contents were heated at 100 °C for 6.5 h. The crude material was purified by silica gel chromatography using a gradient of 15% acetonitrile:methanol (7:1) in dichloromethane providing the desired compound, reference compound I-502 (54 mg, 90% yield) as a solid. white.

1H-NMR (500 MHz, DMSO-d6) 89.10 (d, 1 H), 8.18 (d, 1 H), 8.00 (s, 1 H), 7.41 (s, 1 H) , 7.33 (ap. q, 1 H), 7.24-7.20 (m, 1 H), 7.22 (d, 1 H), 7.11 (ap. t, 1 H), 6 .89 (ap. t, 1 H), 5.86 (s, 2 H), 4.89 (t, 1 H), 3.63 (d, 2 H), 0.85 (m, 2 H) , 0.77 (m, 2H).

Reference compound I-581

The title compound was prepared following general procedure B, except that 3-aminopropanamide (as the HCl salt) was the amine reagent, and the contents were heated at 100 °C for 21 h. The contents were cooled to room temperature, diluted with water and acidified to a pH of 4 with a 1N HCl solution. The resulting precipitate was filtered and dried under vacuum to provide the desired compound, reference compound I-581 (66 mg, 89% yield) as a white solid.

1H-NMR (500 MHz, DMSO-d6) 8 ppm 9.08 (d, 1 H), 8.17 (d, 1 H), 7.65 (t, 1 H), 7.50 (s, 1 H), 7.34 (br. s, 1 H), 7.32 (ap. q, 1 H), 7.24-7.18 (m, 1 H), 7.21 (d, 1 H) , 7.10 (ap. t, 1 H), 6.86 (br. s, 1 H), 6.84 (m, 1 H), 5.90 (s, 2 H), 3.67 (dt , 2H), 2.45 (t, 2H).

Reference compound I-515

A solution of reference compound I-358 in dichloromethane was treated with N,N-diisopropylethylamine (2 equiv.) followed by 0-(7-azabenzotriazol-1-yl)- -N,N,N',N' hexafluorophosphate -tetramethyluronium (HATU, 1.5 equiv.). After 1 h, ammonia (0.5N solution in dioxane, 3 equiv.) was added and the light brownish orange solution was stirred for 21 h. The resulting light tan suspension was diluted with water and extracted with dichloromethane. The combined organic phases were dried over sodium sulfate, filtered, and the solvent was removed in vacuo. The crude material was purified by silica gel chromatography with a gradient of 10-15% acetonitrile:methanol (7:1) in dichloromethane providing the desired compound, reference compound I-515 (36 mg, 73% yield) as a white solid.

1H-NMR (500 MHz, CD3OD) 8 ppm 8.76 (s, 1 H), 8.10 (d, 1 H), 7.42 (s, 1 H), 7.28 (ap. q, 1 H), 7.10 (m, 1 H), 7.05 (ap. t, 1 H), 6.90 (s, 1 H), 6.89 (m, 1 H), 5.96 (s , 2 H), 3.88 (s, 2 H), 1.20 (m, 2 H), 1.02 (m, 2 H).

Reference compound I-536

A solution of reference compound I-86 in dichloromethane was treated with N,N-diisopropylethylamine (2 equiv.) followed by HATU (1.5 equiv.). After 1 h, ammonia (0.5 N solution in dioxane, 3 equiv.) was added and the reaction was stirred for 24 h. The resulting mixture was diluted with water and extracted with dichloromethane. The combined organic phases were dried over sodium sulfate, filtered, and the solvent was removed in vacuo. The crude material was purified by silica gel chromatography with a gradient of 10-25% acetonitrile-methanol (7:1) in dichloromethane providing the desired compound, compound I-536 (33 mg, 81% yield) as a solid. whitish.

1H-NMR (500 MHz, CDCla) 8 ppm 8.47 (d, 1 H), 8.15 (d, 1 H), 7.28 (s, 1 H), 7.22 (ap. q, 1 H), 7.03 (ap. t, 1 H), 6.99 (ap. t, 1 H), 6.94 (ap. t, 1 H), 6.61 (d, 1 H), 6 .56 (br. s, 1 H), 5.99 (d, 1 H), 5.89 (d, 1 H), 5.60 (d, 1 H), 5.50 (br. s, 1 H), 4.57 (ap. t, 1H), 2.32 (m, 1 H), 1.09 (d, 3 H), 1.07 (d, 3 H).

Reference compound I-537

A solution of reference compound I-69 in dichloromethane was treated with N,N-diisopropylethylamine (2 equiv.) followed by HATU (1.5 equiv.). After 1 h, ammonia (0.5 N solution in dioxane, 3 equiv.) was added and the reaction was stirred for 24 h. The resulting mixture was diluted with water and extracted with dichloromethane. The combined organic phases were dried over sodium sulfate, filtered, and the solvent was removed in vacuo. The crude material was purified by silica gel chromatography with a gradient of 10-25% acetonitrile:methanol (7:1) in dichloromethane providing the desired compound, reference compound I-537 (27 mg, 65% yield) as a white solid.

1 H - R M N ( 500 MHz , D M S O - d a ) 5 pp m 9 , 11 ( d , 1 H ) , 8 , 23 ( d , 1 H ) , 7 , 57 ( b r . s , 1 H ) , 7 , 53 (s, 1H), 7, 38 - 7, 30 (m, 2H), 7, 24 - 7, 16 (m, 2H), 7, 16 (d, 1H), 7, 10 (a p .t, 1H), 6, 85 (ap. t, 1H), 5, 87 (s, 2H), 4, 42 (ap. t, 1H), 2, 20 (m, 1H) , 0 , 97 ( a p . t, 6 H ) .

Reference compound I-538

A solution of reference compound I-85 in dichloromethane was treated with W,W-diisopropylethylamine (2 equiv.) followed by HATU (1.5 equiv.). After 1 h, ammonia (0.5 N solution in dioxane, 3 equiv.) was added and the reaction was stirred for 24 h. The resulting mixture was diluted with water and extracted with dichloromethane. The combined organic phases were dried over sodium sulfate, filtered, and the solvent was removed in vacuo. The crude material was purified by silica gel chromatography with a gradient of 10-25% acetonitrile:methanol (7:1) in dichloromethane providing the desired compound, reference compound I-538 (36 mg, 86% yield) as a white solid.

1H-NMR (500 MHz, DMSO-d6) 5 ppm 9.10 (d, 1 H), 8.22 (m, 2 H), 7.40 (br. s, 1 H), 7.36-7 .28 (m, 2 H), 7.23 (m, 1 H), 7.15 (s, 1 H), 7.11 (ap. t, 1 H), 6.98 (br. s, 1 H), 6.86 (ap. t, 1 H), 5.86 (s, 2 H), 1.42 (m, 2 H), 1.02 (m, 2 H).

Reference compound I-546

A suspension of reference compound I-67 in dichloromethane was treated with di(1H-imidazol-1-yl)methanone (CDI, 3 equiv.) and the resulting mixture was heated at 45 °C for 1 h 40 min. After cooling to room temperature, methanesulfonamide (5 equiv.) and 1,8-diazabicyclo[5.4.0]undec-7-ene (DBU, 1 equiv.) were added and the reaction was heated at 45 °C for 1 h. . The resulting mixture was cooled to room temperature, quenched with 1N HCl solution and extracted with dichloromethane/isopropanol (4:1). The crude solid was dissolved in water with the help of 1N NaOH solution and acidified to a pH of 3-4 by adding 1N HCl dropwise. The resulting precipitate was filtered and dried under vacuum to provide the desired compound, reference compound I-546 (39 mg, 80% yield) as a tan solid.

1H-NMR (500 MHz, DMSO-d6) 5 ppm 12.1 (s, 1 H), 9.11 (d, 1 H), 8.27 (d, 1 H), 8.17 (br. s , 1 H), 7.52 (s, 1 H), 7.32 (ap. q, 1 H), 7.22 (m, 1 H), 7.12 (d, 1 H), 7.10 (m, 1 H), 6.79 (ap. t, 1 H), 5.88 (s, 2 H), 4.12 (d, 2 H), 3.17 (s, 3 H).

Reference compound I-566

The title compound was synthesized in 4 steps:

Stage 1: Synthesis of 2-((4-methoxybenzyl)oxy)acetic acid

To a solution of (4-methoxyphenyl)methanol (1 equiv.) and 2-bromoacetic acid (1.2 equiv.) in anhydrous THF at 0 °C was added sodium hydride (60% w/w in mineral oil, 3 equiv.) in 3 parts. The mixture was stirred at 70 °C for 4 h. After cooling to room temperature, water was added and the resulting mixture was washed with hexanes. The aqueous phase was acidified to pH 2 with 1N HCl and extracted with ethyl acetate. The organic layer was dried, filtered and the solvent was removed in vacuo. Purification by silica gel chromatography with 50% ethyl acetate in hexanes provided 2-((4-methoxybenzyl)oxy)acetic acid (0.51 g, 71% yield) as a clear solid.

1H NMR (500 MHz, CDCla) 5 ppm 7.29 (m, 2 H), 6.90 (m, 2 H), 4.59 (s, 2 H), 4.10 (s, 2 H), 3.81 (s, 3H).

Step 2: Synthesis of 2-((4-methoxybenzyl)oxy)-W -(2,2,2-trifluoroethyl)acetamide

A solution of 2-((4-methoxybenzyl)oxy)acetic acid (1 equiv.) in dichloromethane was treated with W,A/-diisopropylethylamine (1.5 equiv.) followed by HATU (1.2 equiv.). After 30 min, W,W-diisopropylethylamine (2 equiv.) and 2,2,2-trifluoroethanamine hydrochloride (2 equiv.) were added and the reaction was stirred for 17 h. The resulting mixture was diluted with water and extracted with dichloromethane. The combined organic phases were dried over sodium sulfate, filtered, and the solvent was removed in vacuo. The crude material was purified by silica gel chromatography using a gradient of 10-20% ethyl acetate/hexanes providing 2-((4-methoxybenzyl)oxy)-A/-(2,2,2-trifluoroethyl)acetamide ( 0.28 g, yield 78%) as a white solid.

1 H R M N (500 M H z, C D C l a) 5 p p m 7, 26 (d, 2 H), 6, 91 (d, 2 H), 6, 89 (b r . s, 1 H), 4, 52 (s, 2H), 4, 02 (s, 2H), 3, 93 (dq, 2H), 3, 82 (s, 3H).

Stage 3: Synthesis of W-(5-fluoro-2-(1-(2-fluorobenzyl)-5-(¡soxazol-3-¡l)-1H-pyrazol-3-¡l)p¡ r¡m¡d¡n-4-¡l)-2-((4-methoxy¡benz¡l)ox¡)-A/-(2,2,2-trifluoroethyl)acetam¡de

A suspension of 3-(3-(4-chloro-5-fluorop¡r¡m¡d¡n-2-¡l)-1-(2-fluorobenzyl)-1H-pyrazole-5 -¡l)¡soxazole (1 equiv.), 2-((4-methoxy¡benz¡l)ox¡)—W-(2,2,2-trifluoroethyl)acetam¡de (1 eq.) and cesium carbonate (0.8 eq.) in anhydrous dioxane was heated at 100 °C for 4 d. The resulting mixture was poured into half-saturated sodium bicarbonate solution and extracted with ethyl acetate. The organic phase was dried over sodium sulfate, filtered and the solvent was removed in vacuo. The crude material was purified by silica gel chromatography with a gradient of 10-20% ethyl acetate/hexanes providing A/-(5-fluoro-2 -(1-(2-fluorobenzyl)-5-(¡soxazol-3-¡l)-1H-pyrazol-3-¡l)pyr¡m¡d¡n-4-¡l) -2-((4-methoxy¡benz¡l)ox¡)-A/-(2,2,2-trifluoroethyl)acetamide (16 mg, 12% yield) as a transparent oil.

1H NMR (500 MHz, CDCla) 8 ppm 8.65 (d, 1 H), 8.49 (d, 1 H), 7.32 (s, 1 H), 7.22 (ap. q, 1 H ), 7.07-6.96 (m, 4 H), 6.89 (ap. t, 1 H), 6.73 (d, 2 H), 6.60 (d, 1 H), 6, 01 (s, 2 H), 4.68 (q, 2 H), 4.33 (s, 2 H), 4.31 (s, 2 H), 3.71 (s, 3 H).

Stage 4: Syntheses of the reference compound I-566

A biphasic solution of W-(5-fluoro-2-(1-(2-fluorobenzyl)-5-(soxazol-3-l)-1H-pyrazole-3- ¡l)- -p¡r¡m¡d¡n-4-¡l)-2-((4-methoxy¡benz¡l)ox¡)-A/-(2,2,2-trifluoroet ¡l)acetam¡de (1 equiv.) in dichloromethane/water (10:1) was treated with 2,3-dichloro-5,6-d¡cyano-p-benzoquinone (DDQ, 1.2 equiv.) and stirred for 20 h. Additional amounts of DDQ (2.4 equiv.) were added and the reaction was stirred for 5 d. The reaction mixture was diluted with dichloromethane and filtered. The crude solution was dried over sodium sulfate, filtered, and the solvent was removed in vacuo. The crude material was purified by silica gel chromatography with a gradient of 15-50% ethyl acetate/hexanes providing the desired compound, the reference compound. to I-566 (2.6 mg, yield 45%) as a white solid.

1H-NMR (500 MHz, CD3OD) 8 ppm 8.78 (d, 1 H), 8.55 (d, 1 H), 7.44 (s, 1 H), 7.26 (ap. q, 1 H), 7.09 (m, 1 H), 7.03 (ap. t, 1 H), 6.87 (d, 1 H), 6.80 (ap. t, 1 H), 5.97 (s, 2 H), 5.16 (s, 2 H), 3.94 (q, 2 H).

Reference compound I-457

The title compound was synthesized in 2 steps:

Stage 1: Syntheses of 3-(4-chloro-5-fluorop¡r¡m¡d¡n-2-¡l)-1-(2-fluorobenzyl)-1H-pyrazole-5- carbon

A suspension of 3-(5-fluoro-6-oxo-1,6-d¡hydro¡r¡m¡d¡n-2-¡l)- -1-(2-fluorobenzyl) -1H-pyrazol-5-carbonatrilo (this compound was described in the previous patent application publication WO2013/101830) in trichloride Phosphoryl (50 equiv.) as solvent was heated at 65 °C for 2 h 15 min. The reaction mixture was blow dried under a stream of nitrogen and then concentrated twice from toluene. The resulting reddish brown oil/solid was dried under vacuum and used in the next step without further manipulation.

Stage 2: Syntheses of the reference compound I-457

The title compound was prepared following general procedure B, except that 3-(4-chloro-5-fluoroprímidin-2-¡l)-1- (2-fluorobenzyl)-1H-pyrazol-5-carbon¡trilo was the reactant of chloroprámide and the acid (2R,3S)-3-met lPeridin-2-carboxylic acid was the amine reagent, and the content was heated at 100 °C for 18 h. The content was cooled to room temperature, diluted with water, acidified to pH 3 with 1N HCl solution and extracted with dichloromethane. The combined organic phases were dried over sodium sulfate, filtered, and the solvent was removed in vacuo. The crude material was purified by reverse phase HPLC using a gradient of 25-80% acetontrile/water (with 0.1% TFA) providing ionizing the desired compound, reference compound I-457 (18 mg, 38% yield over 2 steps) as a white solid.

1H-NMR (500 MHz, CD3OD) 8 ppm 8.32 (d, 1 H), 7.65 (s, 1 H), 7.41 (ap. q, 1 H), 7.36 (ap. t , 1 H), 7.23-7.13 (m, 2 H), 5.71 (s, 2 H), 5.17 (d, 1 H), 4.48 (br. d, 1 H) , 3.78 (ap. t, 1 H), 2.13 (m, 1 H), 1.94 (m, 1 H), 1.78 (m, 2 H), 1.54 (m, 1 H), 1.22 (d, 3H).

Reference compound I-474

The title compound was prepared according to general procedure B, except that 3-(4-chloro-5-fluoropyrmidin-2-1)-1 was used. -(2-fluorobenzyl)-1H-pyrazol-5-carbonitrile (generated in step 1 for the synthesis of the reference compound I-457 instead of the intermediate product 1, 2-(aminomethyl)-1,1,1,3,3,3-hexafluoropropan-2-ol was the amine reagent and the content was heated to 100 °C for 16 h. The content was cooled to room temperature, diluted with water, acidified to pH 3 with 1N HCl solution and extracted with dichloromethane. The organic phases were dried over sodium sulfate, filtered, and the solvent was removed in vacuo. The crude material was purified by phase HPLC. converse using a gradient of 25-80% acetonitril/water (with 0.1% TFA) providing the desired compound, the reference compound I-474 (31 mg , yield 49 % for 2 stages from 3-(5-fluoro-6-oxo-1,6-dihydro¡r¡m¡d¡n-2-¡l) -1-

- (2 - f l u o ro o b e n c ¡ l ) - 1 H - p¡ r a z o l - 5 - c a r b o n i t l o ) as a d o b l a n c o s o l d .

Reference compound I-480

The title compound was prepared following general procedure B, except that 3-(4-chloro-5-fluoropyrimidin-2-yl)-1-(2-fluorobenzyl)-1H-pyrazole-5-carbonitrile (generated in step 1 for the synthesis of reference compound I-457 instead of intermediate 1, and 1-(1-carboxycyclopropyl)-A/-methylmethanamine (as the HCl salt) was the amine reagent and the content was heated at 100 °C for 17 h. The content was cooled to room temperature, diluted with water, acidified to pH 3 with 1N HCl solution and extracted with dichloromethane. The combined organic phases were dried over sodium sulfate. Filtered, and the solvent was removed in vacuo. The crude material was purified by preparative HPLC using a gradient of 15-70% acetonitrile/water (with 0.1% TFA) to provide the desired compound, reference compound I-480. (90 mg, 66% yield over 2 steps from 3-(5-fluoro-6-oxo-1,6-dihydropyrimidin-2-yl)-1-(2-fluorobenzyl)-1H-pyrazole-5-carbonitrile ) as a whitish solid.

1H-NMR (500 MHz, DMSO-d6) 5 ppm 12.3 (br. s, 1 H), 8.24 (d, 1 H), 7.66 (s, 1 H), 7.44 (ap .q, 1 H), 7.36 (ap. t, 1 H), 7.30-7.22 (m, 2 H), 5.65 (s, 2 H), 3.99 (s, 2 H), 3.24 (d, 3 H), 1.13 (m, 2 H), 1.01 (m, 2 H).

Reference compound I-476

A solution of reference compound I-474 in 1 N NaOH solution (excess) was heated at 65 °C for 70 min. The reaction mixture was cooled to room temperature and acidified to a pH of 3 with a 1N HCl solution. The resulting precipitate was filtered and dried under vacuum to provide the desired compound, reference compound I-476 (13 mg, 77% yield) as a white solid.

1H-NMR (500 MHz, DMSO-d6) 5 ppm 8.50 (s, 1 H), 8.34 (d, 1 H), 8.03 (br. t, 1 H), 7.35 (ap .q, 1 H), 7.32 (s, 1 H), 7.22 (m, 1 H), 7.13 (ap. t, 1 H), 7.02 (ap. t, 1 H) , 5.87 (s, 2 H), 4.13 (d, 2 H). The exchangeable carboxylic acid proton was not observed.

Reference compound I-481

A suspension of reference compound I-480 in water was treated with 1 N NaOH solution (2 equiv.) and stirred at room temperature for 18 h. The reaction mixture was acidified to pH 3 with a 1N HCl solution. The resulting precipitate was filtered and dried under vacuum to provide the desired compound, reference compound I-481 (7.4 mg, 64% yield) as a white solid.

1H-NMR (500 MHz, DMSO-d6) 5 ppm 12.3 (br. s, 1 H), 8.22 (d, 1 H), 8.14 (br. s, 1 H), 7.60 (br. s, 1 H), 7.53 (s, 1 H), 7.33 (ap. q, 1 H), 7.21 (m, 1 H), 7.12 (ap. t, 1 H), 6.90 (ap. t, 1 H), 5.90 (s, 2 H), 3.99 (s, 2 H), 3.24 (d, 3 H), 1.14 (m , 2H), 1.02 (m, 2H).

Reference compound I-327

To a mixture containing 1,2-diethoxycyclobutenedione (1.3 equiv.) and sodium hydride [60% dispersion in mineral oil] (1 equiv.) in THF was added intermediate 2 (1 equiv.). The mixture was stirred at 0 °C for 1 h and then removed from the ice bath and allowed to stir at 23 °C for 24 h. The mixture was diluted with ethyl acetate and washed with 1N HCl solution. The organic layer was dried over MgSO4, filtered and evaporated to give a solid. The crude material was purified by silica gel chromatography with a gradient of 0-100% ethyl acetate/hexanes providing the desired compound, reference compound I-327 (90 mg, 43% yield) as a light yellow solid.

1H NMR (400 MHz, DMSO-d6) 5 ppm 9.12 (d, 1 H), 8.69 (d, 1 H), 7.67 (s, 1 H), 7.30 - 7.38 ( m, 1 H), 7.29 (d, 1 H), 7.24 (d, 1 H), 7.20 (d, 1 H), 7.09-7.15 (m, 1 H), 6 .86-6.92 (m, 1 H), 5.92 (s, 2 H), 4.81 (q, 2 H), 1.36 (t, 3 H).

Reference compound I-402

A mixture of reference compound I-327 (1 equiv.) and HCl [1.0 M aqueous solution] (1 equiv.) in MeOH was heated at 65 °C for 2 h. Upon cooling the mixture to 23 °C, a yellow precipitate formed which was collected by filtration and clarified with a minimal amount of methanol. The collected precipitate was dried under vacuum to provide the desired compound, reference compound I-402 (50 mg, 76% yield) as a yellow solid.1

1H NMR (500 MHz, DMSO-d6) 5 ppm 9.08 - 9.17 (m, 1 H), 8.64 (d, 1 H), 7.73 (s, 1 H), 7.30 - 7.42 (m, 1 H), 7.28 (s, 1 H), 7.17 - 7.26 (m, 2 H), 7.12 (t, 1 H), 6.90 - 7, 04 (m, 1 H), 5.85 - 6.03 (m, 2 H).

Reference compound I-456

To a cold solution of triethylamine (1.5 equiv.) in dichloromethane at 0 °C was added chlorosulfonyl isocyanate (1.5 equiv.). The mixture was stirred at 0 °C for 30 min. Intermediate 2 (1 equiv.) ytert-butanol (1.5 equiv.) was added to this mixture, and the content was stirred at 23 °C for 24 h. The mixture was diluted with ethyl acetate and washed with water. The precipitate was removed by filtration. The organic layer was dried over MgSO4, filtered and concentrated in vacuo to provide a crude oil, which was purified by silica gel chromatography with a gradient of 0-100% ethyl acetate/hexanes providing N-(2-(1 Fed-butyl -(2-fluorobenzyl)-5-(isoxazol-3-yl)-1H-pyrazol-3-yl)pyrimidin-4-yl)sulfamoylcarbamate, the desired Boc-protected sulfonamide intermediate. This intermediate was dissolved in methanol and treated with HCl [a 4.0 M solution in 1,4-dioxane] (5 equiv.) and stirred at 23 °C for 24 h. The mixture was concentrated in vacuo to provide the desired compound, reference compound I-456 (26 mg, 6% yield, HCl salt) as a white solid.

1H NMR (500 MHz, CD3OD) 8 ppm 8.85 (d, 1 H), 8.54 (d, 1 H), 7.88 (s, 1 H), 7.26 - 7.34 (m, 2 H), 7.00 - 7.14 (m, 4 H), 6.05-6.08 (m, 2 H).

Reference compound I-467

The title compound was synthesized in 3 steps:

Stage 1: Synthesis of 2-(bromomethyl)-3,3,3-trifluoro-2-hydroxypropanoic acid.

A mixture of 2-(bromomethyl)-3,3,3-trifluoro-2-hydroxypropanenitrile (1 equiv.), water (1 equiv.), and concentrated sulfuric acid (4 equiv.) was heated to 110 °C in a vial. sealed for 1 hour. The mixture was poured onto ice and extracted with diethyl ether. The organic layer was dried over MgSO4, filtered and concentrated in vacuo to provide 2-(bromomethyl)-3,3,3-trifluoro-2-hydroxypropanoic acid (1.3 g, 33% yield) as a clear oil. 1H NMR (500 MHz, CDCla) 8 ppm 3.89 (d, 1 H), 3.63 - 3.69 (m, 1 H).

Stage 2: Synthesis of 2-(aminomethyl)-3,3,3-trifluoro-2-hydroxypropanoic acid.

A mixture of ammonium hydroxide [28% solution in water] (10 equiv.) and 2-(bromomethyl)-3,3,3-trifluoro-2-hydroxypropanoic acid (1 equiv.) was stirred at 23 °C for 24 hours The mixture was concentrated in vacuo. The resulting solid was treated with a minimum amount of ethanol. The precipitate was collected by filtration and dried under vacuum to provide 2-(aminomethyl)-3,3,3-trifluoro-2-hydroxypropanoic acid (412 mg, 43% yield) as a white solid.

1H NMR (500 MHz, DMSO-d6) 8 ppm 2.86 - 3.27 (m, 2 H).

Stage 3: Synthesis of reference compound I-467

The title compound was prepared following general procedure B, except that 2-(aminomethyl)-3,3,3-trifluoro-2-hydroxypropanoic acid (4 equiv.) was the amine reagent, 6 equiv. of triethylamine was used. , and the content was heated to 85 °C as a solution in 1,4-dioxane/water (4:1) for 24 h. The mixture was cooled to 23°C and diluted with ethyl acetate. The organic layer was washed with saturated ammonium chloride solution, dried over MgSO4, filtered and concentrated in vacuo to give a crude solid. The crude material was purified by silica gel chromatography with a gradient of 0-100% ethyl acetate/hexanes providing the desired compound, reference compound I-467 (50 mg, 7% yield for step 3) as a solid white.1

1H NMR (500 MHz, DMSO-d6) 8 ppm 8.28 (d, 1 H), 7.59 (t, 1 H), 7.46 (s, 1 H), 7.30 - 7.36 ( m, 1 H), 7.16 - 7.24 (m, 2 H), 7.10 (t, 1 H), 6.91 (t, 1 H), 5.88 (s, 2 H), 4.24 (dd, 1 H), 3.84 (dd, 1 H).

Reference compound I-468

A mixture of CDI (6 equiv.) and 3,3,3-trifluoro-2-hydroxy-2-(trifluoromethyl)propanoic acid (6 equiv.) in THF was heated at 90 °C for 1 hour. To this mixture, 2-(1-(2-fluorobenzyl)-5-(isoxazol-3-yl)-1H-pyrazol-3-yl)-5-morpholinopyrimidin-4-amine was added (this intermediate was described in previously published patent application WO2012/3405A1; 1 equiv.). The mixture was stirred at 90 °C for 24 h. The mixture was diluted in ethyl acetate and washed with 1N HCl solution. The organic layer was dried over MgSO4, filtered and evaporated to give a crude oil. The oil was purified by silica gel chromatography with a gradient of 0-100% ethyl acetate/hexanes providing the desired compound, reference compound I-468 (18 mg, 62% yield) as a light yellow solid.

1 H R M N (500 M H z, C D 3O D) 8 p p m 8, 76 (d, 1 H), 8, 64 (s, 1 H), 7, 60 - 7, 63 (m, 1 H), 7, 20 - 7, 26 (m, 1 H), 7, 00 - 7, 06 (m, 1 H), 6, 98 (t, 1 H), 6, 92 (d, 1 H), 6, 74 - 6 , 83 ( m , 1 H ) , 5 , 93 ( s , 2 H ) , 3 , 88 - 3 , 92 ( m , 4 H ) , 3 , 04 - 3 , 09 ( m , 4 H ) .

REFERENCE COMPOSITION I - 473

To a mixture of 5-fluoro-2-(1-(2-fluorobenzyl)-5-(isoxazol-3-yl)-1H-pyrazol-3-yl)- -pyrimidin-4-amine (this intermediate product has been previously described in published patent application WO2012/3405A1, 1 equiv.) and 3,3,3-trifluoro-2-(trifluoromethyl)-2- -((trimethylsilyl)oxy)propanoyl chloride (3 equiv.) [prepared according to the procedure described in Aicher, T.D.et al. J. Med. Chem.2000,43,245, Method J.] in THF at 23 °C, LiHMDS (2.0 M in THF, 3 equiv.) was added very slowly. The exothermic reaction turned dark brown immediately. The mixture was stirred at 23 °C for 1 h, then diluted in ethyl acetate and washed with 1N HCl solution. The precipitate was removed by filtration. The organic layer was dried over MgSO4, filtered, and concentrated in vacuo to provide a crude oil. The oil was purified by silica gel chromatography with a gradient of 0-30% ethyl acetate/hexanes to provide the desired compound, reference compound I-473 (11 mg, 3% yield) as a yellow solid.

1H NMR (500 MHz, CD3OD) 8 ppm 8.83 (d, 1 H), 8.80 (s, 1 H), 7.60 (s, 1 H), 7.27 - 7.33 (m, 1 H), 7.09 - 7.15 (m, 1 H), 7.06 (t, 1 H), 6.94 (d, 1 H), 6.90 (t, 1 H), 6.00 ( s, 2 H).

Reference compound I-477

To a mixture of 5-fluoro-2-(1-(2-fluorobenzyl)-5-(isoxazol-3-yl)-1H-pyrazol-3-yl)- -pyrimidin-4-amine (described in WO2012/3405 A1, 1 equiv.) and morpholine-4-carbonyl chloride (1.2 equiv.) in THF at 23 °C, LiHMDS (2.0 M in THF, 1.2 equiv.) was added, very slowly. The mixture was stirred at 23 °C for 24 h. The mixture was diluted in ethyl acetate and washed with 1N HCl solution. The organic layer was dried over MgSO4, filtered and evaporated to give a crude oil. The oil was purified by silica gel chromatography using a 0-5% methanol/DCM gradient providing the desired product, reference compound I-477 (24 mg, 18% yield) as a light yellow solid.

1H NMR (500 MHz, CD3OD) 8 ppm 8.49 - 8.54 (m, 1 H), 7.47 (s, 1 H), 7.33 - 7.42 (m, 1 H), 7, 23 - 7.30 (m, 1 H), 7.12 -7.22 (m, 1 H), 6.99 - 7.11 (m, 2 H), 6.87 (d, 1 H), 5.95 (s, 2 H), 3.72 (q, 4 H), 3.56 - 3.62 (m, 4 H).

Reference compound I-482

To a cold mixture of triphosgene (0.75 equiv.) and 3-bromo-1,1,1-trifluoro-propan-2-ol (1.5 equiv.) in dichloromethane was added pyridine (1.5 equiv.) . The mixture was stirred at 0 °C for 30 min. In a separate flask, a suspension of intermediate 2 (1 equiv.) in pyridine was cooled to 0 °C. This suspension was transferred to the mixture of triphosgene and bromopropanol using a syringe. The resulting mixture was heated at 60 °C for 24 h. The contents were diluted in ethyl acetate and washed with 1N HCl solution. The organic layer was dried over MgSO4, filtered and evaporated to give a crude oil. The oil was purified by silica gel chromatography with a gradient of 0-100% ethyl acetate/hexanes providing the desired compound, reference compound I-482 (46 mg, 9% yield) as a white solid.

1H NMR (500 MHz, CDCh) 8 ppm 8.79 (d, 1 H), 8.50 (d, 1 H), 8.11 -8.14 (m , 1 H), 7.43 (s, 1 H), 7.19-7.27 (m, 1 H), 6.96 - 7.10 (m, 2 H), 6.87 - 6.93 (m, 1 H), 6.62 ( d, 1 H), 5.99 - 6.03 (m, 2 H), 4.99 - 5.07 (m, 1 H), 4.57 - 4.65 (m, 1 H), 4, 49-4.56 (m, 1H).

Reference compound I-492

A solution of 2,2-bis(trifluoromethyl)-2-hydroxyacetic acid (3 equiv.) in THF was heated to 80 °C for 1 h. To this mixture was added a solution of 5-fluoro-2-(1-(2-fluorobenzyl)-5-(isoxazol-3-yl)-1H-pyrazol-3-yl)pyrimidin-4-amine (intermediate product described in document WO2012/3405 A1 above; 1 equiv.) in NMP. The resulting mixture was heated in a microwave at 200 °C for 1 h. The contents were diluted in ethyl acetate and washed with water. The organic layer was dried over MgSO4, filtered and dried in vacuo to give a crude oil. The oil was purified by silica gel chromatography with a gradient of 0-100% ethyl acetate/hexanes providing the desired product, reference compound I-492 (25 mg, 8% yield) as a yellow solid.

1H NMR (500 MHz, CDCh) 8 ppm 8.48 (d, 1 H), 7.37 (s, 1 H), 7.18 - 7.25 (m, 2 H), 7.00 - 7, 06 (m, 1 H), 6.98 (t, 1 H), 6.85 (t, 1 H), 6.59 (d, 1 H), 5.99 (s, 2 H).

Reference compound I-493

A mixture of reference compound I-403 (1 equiv.), HOBT (3 equiv.), triethylamine (3 equiv.), HATU (3 equiv.) and cyclopropylamine (3 equiv.) in DMF was stirred at 23 °C for 24 hours. The mixture was diluted with ethyl acetate and washed sequentially with 1N HCl solution, saturated sodium bicarbonate solution and brine. The organic layer was dried over MgSO4, filtered and concentrated in vacuo to give a crude oil. The oil was purified by silica gel chromatography with a gradient of 0-100% ethyl acetate/hexanes providing the desired product, reference compound I-493 (20.4 mg, 27% yield) as a white solid. 1

1H NMR (500 MHz, CD3OD) 8 ppm 8.76 - 8.80 (m, 1 H), 8.25 - 8.29 (m, 1 H), 7.47 - 7.49 (m, 1 H) ), 7.24 - 7.31 (m, 1 H), 7.07 - 7.14 (m, 1 H), 7.03 (t, 1 H), 6.87 - 6.90 (m, 1 H ), 6.77 (t, 1 H), 5.95 - 5.99 (m, 2 H), 5.87 - 5.94 (m, 1 H), 2.70 -2.77 (m, 1H), 0.70 - 0.78 (m, 2H), 0.47 - 0.54 (m, 2H).

Reference compound I-504

The title compound was synthesized in 2 steps:

Step 1: Synthesis of 2-bromo-3,3,3-trifluoropropyl (2-(1-(2-fluorobenzyl)-5-(isoxazol-3-yl)-1H-pyrazol-3-yl)pyrimidin-4- il)carbamate.

To a cold mixture of triphosgene (0.9 equiv.) and 2-bromo-3,3,3-trifluoropropan-1-ol (2 equiv.) in THF was added pyridine (2 equiv.). The mixture was stirred at 0 °C for 30 min. In a separate flask, a suspension of intermediate 2 (1 equiv.) in pyridine (2 equiv.) was cooled to 0 °C. The mixture of triphosgene and bromopropanol was added to this suspension through a syringe, and the resulting mixture was heated to 60 °C for 24 h. The contents were concentrated in vacuo and the residue was diluted with ethyl acetate and washed with 1N HCl solution. The organic layer was dried over MgSO4, filtered and concentrated in vacuo to give a crude oil. The oil was purified by silica gel chromatography with a gradient of 0-100% ethyl acetate/hexanes providing the desired carbamate intermediate, 2-bromo-3,3,3-trifluoropropyl (2-(1-(2 -fluorobenzyl)-5-(isoxazol-3-yl)-1H-pyrazol-3-yl)pyrimidin-4-yl)carbamate (77 mg, 4% yield) as a light brown oil.

1H NMR (500 MHz, CDCh) 8 ppm 8.46 - 8.48 (m, 1 H), 8.07 (br. s., 1 H), 7.86 (d, 1 H), 7.44 - 7.47 (m, 1 H), 7.17 - 7.24 (m, 1 H), 7.00 - 7.07 (m, 1 H), 6.93 - 7.00 (m, 1 H), 6.78 - 6.85 (m, 1 H), 6.57 - 6.62 (m, 1 H), 6.02 (s, 2 H), 4.28 (quint, 1 H) , 3.93-4.15 (m, 2 H).

Stage 2: Synthesis of reference compound I-504

To a solution of 2-bromo-3,3,3-trifluoropropyl (2-(1-(2-fluorobenzyl)-5-(isoxazol-3-yl)-1H-pyrazol-3-yl)pyrimidin-4-yl )carbamate in THF LiHMDS (2.0 M in THF, 1 equiv.) was added. The mixture was sealed and heated at 60 °C for 2 days. The mixture was diluted with ethyl acetate and washed with water. The organic layer was dried over MgSO4, filtered and evaporated to give a crude oil. The oil was purified by silica gel chromatography using a gradient of 0-100% ethyl acetate/hexanes and recrystallized from a mixture of diethyl ether hexanes providing the desired product, reference compound I-504 (7 mg, yield 11%) as a white solid.

1H NMR (500 MHz, DMSO-d6) 8 ppm 9.13 (s, 1 H), 8.84 (s, 1 H), 7.99 (d, 1 H), 7.70 (s, 1 H) ), 7.31 - 7.38 (m, 1 H), 7.19 - 7.26 (m, 2 H), 7.12 (t, 1 H), 6.91 (t, 1 H), 6.03 - 6.09 (m, 1 H), 5.86 - 5.98 (m, 2 H), 4.73 - 4.80 (m, 2 H).

Reference compound I-544

A mixture of reference compound I-419 (1 equiv.) and lithium aluminum hydride (2 equiv.) in THF was heated at 60 °C for 24 h. The mixture was cooled to 23 °C, then treated sequentially with water (x ml/x g lithium aluminum hydride), 15% NaOH (aq) (x ml/x g lithium aluminum hydride), and water (3x ml/x g lithium aluminum hydride). The precipitate was removed by filtration and the filtrate was concentrated in vacuo to provide the intermediate amine as a yellow solid. The intermediate product was suspended in THF, and a solution of methanesulfonyl chloride (1 M in THF, 2 equiv.) and pyridine (3 equiv.) in THF was added to the suspension dropwise via syringe. The mixture was stirred at 23 °C for 3 h, then diluted in ethyl acetate and washed with 1N HCl solution. The organic layer was dried, filtered and evaporated to provide a crude oil. The oil was purified by silica gel chromatography with a gradient of 0-100% ethyl acetate/hexanes providing the desired product, reference compound I-544 (4 mg, 19% yield) as a yellow solid.

1H NMR (400 MHz, CD3OD) 8 ppm 8.71 - 8.78 (m, 1 H), 8.22 (d, 1 H), 7.51 (s, 1 H), 7.22 - 7, 32 (m, 1 H), 6.97 - 7.15 (m, 2 H), 6.89 - 6.96 (m, 1 H), 6.69 - 6.83 (m, 1 H), 5.99 - 6.05 (m, 2 H), 5.56 (s, 1 H), 3.03 - 3.23 (m, 2 H), 2.68 - 2.83 (s, 3 H) ), 1.95-2.08 (m, 2 H).

Reference compound I-575

The title compound was synthesized in 3 steps:

Step 1: Synthesis of (R)-1-((tert-butyldimethylsilyl)oxy)-3-((5-fluoro-2-

-(1-(2-fluorobenzyl)-5-(isoxazol-3-yl)-1H-pyrazol-3-yl)pyrimidin-4-yl)amino)propan-2-ol.

A mixture of reference compound I-316 (1 equiv.), imidazole (2 equiv.) and TBDMS-Cl (1 equiv.) in DMF was stirred at rt for 24 h. The mixture was diluted in ethyl acetate and washed with 1N HCl solution. The organic layer was dried, filtered and evaporated to give a crude oil. The oil was purified by silica gel chromatography with a gradient of 0-100% ethyl acetate/hexanes providing the desired intermediate, (R)-1-((tertbutyldimethylsilyl)oxy)-3- -((5-fluoro -2-(1-(2-fluorobenzyl)-5-(isoxazol-3-yl)-1H-pyrazol-3-yl)pyrimidin-4-yl)amino)propan-2-ol (258 mg, 63% yield ).

1 H R M N (500 M H z, C D C l a) 8 p p m 8, 34 (d, 1 H), 8, 04 - 8, 07 (m, 1 H), 7, 23 (b r. s., 1 H), 7 , 06 - 7 , 13 ( m , 1 H ) , 6 , 82 - 6 , 97 ( m , 3 H ) , 6 , 76 ( t , 1 H ) , 5 , 84 - 5 , 90 ( m , 2 H ) , 3 , 80 - 3 , 89 ( m , 1 H ) , 3 , 47 - 3 , 66 ( m , 4 H ) , 0 , 79 - 0 , 84 ( m , 9 H ) , 0 , 03 ( m , 6 H).

Stage 2: Synthesis of (R)-5-(((tert-but¡ld¡met¡ls¡l¡l)ox¡)met¡l)-3-(5-fluoro- -2-(1-( 2-fluorobenzyl)-5-(soxazol-3-yl)-1H-pyrazol-3-yl)pyrimidin-4-yl)oxazolidin-2-one.

A mixture of (R)-1-((tert-but¡ld¡methyls¡l¡l)ox¡)-3-((5-fluoro- -2-(1-(2-fluorobenzyl) -5-(¡soxazol-3-¡l)-1H-pyrazol-3-¡l)pyr¡m¡d¡n-4-¡l)amino)propan-2-ol (1 equ¡v.), 2,6-dimet¡lprídine (2 equiv.) and triphosgene (0.7 equiv.) in THF stirred at 23° C for 30 min. The mixture was then heated at 60 °C for 24 h. The content was diluted in ethyl acetate and washed with water. The organic layer was dried, filtered and evaporated giving a crude oil. The oil was purified by silica gel chromatography with a gradient of 0-100% ethyl acetate/hexanes providing the protected carbamate intermediate. with desired TBS, the (R)-5-(((tert-but¡ld¡met¡ls¡l¡l)ox¡)met¡l)-3-(5-fluoro-2-(1-( 2-fluorobenzyl)-5-(¡soxazol-3-¡l)-1H-pyrazol-3-¡l)pyr¡m¡n-4-¡l)oxazol¡din -2-one (221 mg, yield 82%).

1H NMR (500 MHz, CDCla) 8 ppm 8.54 (d, 1 H), 8.36 (d, 1 H), 7.24 (s, 1 H), 7.05 - 7.13 (m, 1 H), 6.84 - 6.97 (m, 2 H), 6.72 - 6.81 (m, 1 H), 6.47 (d, 1 H), 5.87 (s, 2 H) ), 4.65 - 4.74 (m, 1 H), 4.24 - 4.32 (m, 1 H), 4.16 (dd, 1 H), 3.83 (m, 2H), 0 .74-0.82 (m, 9 H), 0.00 (d, 6 H).

Stage 3: Syntheses of the reference compound I-575

To a cold solution of (R)-5-(((tert-but¡ld¡met¡ls¡l¡l)ox¡)met¡l)-3-(5-fluoro-2--(1 -(2-fluorobenzyl)-5-(¡soxazol-3-¡l)-1H-pyrazol-3-¡l)pyr¡m¡n-4-¡l)oxazol¡d n-2-one (1 equiv.) in THF at 25 °C a solution of TBAF (1 M in THF, 1 equiv.) was added. After stirring the mixture at 23 °C for 30 min, the mixture was quenched with water and diluted with ethyl acetate. The organic layer was dried, filtered and evaporated giving a crude oil. The oil was purified by silica gel chromatography using a gradient of 0-100% ethyl acetate/hexanes. Further purification by recrystallization from a dichloromethane-diethyl ether mixture gave the desired compound, the reference compound. I-575 (10 mg, 4% yield for 3 steps) as a white solid.

1H NMR (500 MHz, DMSO-d6) 8 ppm 8.25 (d, 1 H), 8.02 (t, 1 H), 7.53 (s, 1 H), 7.30 - 7.39 ( m, 1 H), 7.17 - 7.25 (m, 2 H), 7.11 (td, 1 H), 6.85 - 6.91 (m, 1 H), 5.88 (s, 2 H), 5.06 (dq, 1 H), 4.61 (t, 1 H), 4.45 (dd, 1 H), 3.83 (m, 2 H).

Reference compound I-490

A mixture of 4,4,4-trifluoro-3-hydroxyl-3-(trifluoromethyl)butanoic acid (1.5 equiv.) and CDI (1.5 eq.) in THF was heated to reflux for 2 h. Intermediate product 2 (1 equiv.) was added to this mixture in one part. The mixture was diluted in ethyl acetate and washed with 1N HCl solution. The organic layer was dried, filtered and evaporated giving an oil. The oil was purified by silica gel chromatography using a Socratic gradient of 80% ethyl acetate in hexanes providing the desired product. , the reference compound I-490 (2 mg, yield 1.2%) as a white solid.

1H-NMR (500 MHz, CDCla) 8 ppm 8.80 (d, 1 H), 8.51 (s, 1 H), 8.05 (d, 1 H), 7.44 -7.50 (m , 1 H), 7.19 - 7.32 (m, 2 H), 6.95 - 7.08 (m, 2 H), 6.87 (d, 1 H), 6.58 - 6.65 (m, 1 H), 5.97 (s, 2H), 2.93 - 2.99 (m, 2 H).

Reference compound I-496

The title compound was prepared following general procedure B, except that (5-methyl-1,3,4-oxadilazole-2-l)methanamine (as HCl salt) was the amine reagent, and the content was heated at 110 °C for 24 h as a solution in THF/water (10:1). The contents were cooled to 23 °C, and the organic solvents were removed in vacuo. The resulting residue was purified by reverse phase HPLC providing the desired product, the reference compound I-496 (81 mg, yield 64%) as a solid. do white.

1H NMR (500 MHz, DMSO-d6) 8 ppm 9.09 - 9.12 (m, 1 H), 8.57 (br. s., 1 H), 8.32 (d, 1 H), 7 .48 (s, 1 H), 7.30 - 7.36 (m, 1 H), 7.19-7.25 (m, 1 H), 7.16 (d, 1 H), 7.11 (t, 1 H), 6.84 (t, 1 H), 5.88 (s, 2 H), 4.92 (d, 2 H), 2.45 (s, 3 H).

Reference compound I-508

The title compound was prepared in 3 steps:

Stage 1: Syntheses of fed-butyl (1-(cyclopropylamine)-3-methyl-1-oxobutan-2-l)carbamate

To a solution of 2-((ted-butoxycarbon¡l)amino)-3-methylbutane¡ic acid (1 eq) and cyclopropanamine (1 Eq) in THF (10 ml) PyAOP (1.0 eq) was added followed by DIPEA (3 eq). The reaction mixture was stirred at room temperature for 4 hours. With complete conversion of starting material into the desired product, the solvent was removed in vacuo and purified with flash chromatography eluent with ethyl acetate/ hexane 1:1. Fractions containing the desired product were collected and concentrated to provide the amide intermediate product as an oil.

Stage 2: Synthesis of 2-amino-N-cidopropyl-3-methylbutanamide

The tert-butyl amide (1-(cidopropylamino)-3-methyl-1--oxobutan-2-yl)carbamate intermediate (1 equiv.) was dissolved in dichloromethane and TFA (3:1 ratio) and stirred. for 4 h at 23 °C. The solvent was removed in vacuo providing the free amine intermediate 2-amino-N-cyclopropyl-3-methylbutanamide (0.25 g, 42% yield) as a semisolid.

Stage 3: Synthesis of reference compound I-508

The title compound was prepared following general procedure B, except that 2-amino-N-cyclopropyl-3-methylbutanamide was the amine reagent and the content was heated at 110 °C for 24 h as a solution in THF/water (10:1). The contents were cooled to 23 °C, and the organic solvents were removed in vacuo. The resulting residue was purified by reverse phase HPLC to provide the desired product, reference compound I-508 (15 mg, 22% yield) as a white solid.

1H NMR (500 MHz, CDCh) 8 ppm 8.51 - 8.55 (m, 1H), 8.15 (d, 1H), 7.40 - 7.45 (m, 1H), 7.23 - 7.28 (m, 1 H), 6.99 -7.12 (m, 2 H), 6.70-6.75 (m, 1 H), 6.62-6.69 (m, 1 H), 6.65 (br. s., 1H), 5.93- 5.98 (m, 2 H), 4.58 (t, 1 H), 2.75 (tq, 1 H), 2 .31 (dq, 1 H), 0.99 - 1.08 (m, 6 H), 0.67 - 0.79 (m, 2 H), 0.44 - 0.55 (m, 2 H) .

Reference compound I-509

The title compound was prepared following general procedure B, except that 2-((trifluoromethyl)thio)ethanamine was the amine reagent, and the content was heated at 110 °C for 24 h as a solution in THF/water ( 10:1). The contents were cooled to 23 °C, and the organic solvents were removed in vacuo. The resulting residue was purified using a gradient of 5-50% ethyl acetate in hexane providing the desired product, reference compound I-509 (81 mg, 60% yield) as an off-white solid.

1H NMR (500 MHz, CDCla) 8 ppm 8.45 - 8.48 (m, 1 H), 8.20 (d, 1 H), 7.30 (s, 2 H), 7.16 - 7, 23 (m, 1 H), 7.00 - 7.06 (m, 1 H), 6.97 (t, 1 H), 6.86 (t, 1 H), 6.57 - 6.60 ( m, 1 H), 5.95 - 6.01 (m, 2 H), 3.96 (q, 2 H), 3.27 (t, 2 H).

Reference compound I-514

To a stirred solution of reference compound I-509 (1 equiv.) in dichloromethane was added mCPBA (2 equiv.), and the mixture was stirred for 12 h. The solvent was removed in vacuo and the resulting residue was purified by reverse phase HpLC to provide the desired product, reference compound I-514 (5 mg, 6% yield) as a white solid.

1H NMR (500 MHz, CDCh) 8 ppm 8.41 (d, 1 H), 8.17 (d, 1 H), 7.28-7.32 (m, 1 H), 7.10-7, 17 (m, 1 H), 6.89-6.99 (m, 2 H), 6.80 - 6.86 (m, 1 H), 6.53 (d, 1 H), 5.99 ( d, 1 H), 5.90 (s, 2 H), 4.20 (q, 2 H), 3.70 (t, 2 H).

Reference compound I-529

The title compound was prepared following general procedure B, except that 1,1,1,3,3,3-hexafluoro-2-((methylamino)methyl)propan-2-ol was the amine reagent, and the content was heated at 110 °C for 24 h as a solution in THF/water (10:1). The contents were cooled to 23 °C, and the organic solvents were removed in vacuo. The resulting residue was purified by reverse phase HPLC to provide the desired product, reference compound I-529 (2.5 mg, 2% yield) as an off-white solid.

1H NMR (500 MHz, CDCh) 8 ppm 8.53 (d, 1 H), 8.36 (d, 1 H), 7.42 (br. s., 1 H), 7.23 -7.28 (m, 2 H), 7.03 - 7.25 (m. 2 H), 6.64 (s, 1 H), 5.95 (s, 2 H), 4.22 (br. s., 1 H), 3.49 - 3.53 (m, 3 H), 3.02 - 3.08 br. 2H).

Reference compound I-545

The title compound was prepared following general procedure B, except that (1-(methylsulfonyl)cyclopropyl)methanamine (as the HCl salt) was the amine reagent, and the contents were heated at 110 °C for 24 h as a solution in dioxane/water (10:1). The contents were cooled to 23 °C, and the organic solvents were removed in vacuo. The resulting residue was purified by reverse phase HPLC to provide the desired product, reference compound I-545 (81 mg, 59% yield) as a white solid.

1 H R M N (500 M H z, D M S O - d 6) 8 p p m 9, 12 (d, 1 H), 8, 30 (d, 1 H), 8, 19 (b r. s., 1 H), 7, 55 - 7, 61 (m, 1 H), 7, 31 - 7, 38 (m, 1 H), 7, 19 - 7, 26 (m, 2 H), 7, 13 (t, 1 H), 6 , 96 (t, 1 H), 5, 89 (s, 2 H), 4, 04 (d, 2 H), 3, 09 (s, 3 H), 1, 22 (s, 4 H).

REFERENCE COMPOSITION I - 567

The title compound was prepared in 5 steps:

Step 1: Synthesis of methyl 1-(((ferf-butoxy¡carbon¡l)am¡no)methyl¡cyclopropanecarbox¡late

To a stirred solution of 1-(((ferf-butoxy¡carbon¡l)amino)methyl)cyclopropanecarboxylic acid (1 equiv.) in diethyl ether and methanol (ratio 5:1) was added slowly (diazomethyl)trimethylsilane (1 equiv.) at 25 °C. The mixture was stirred overnight, and the solvents were removed in vacuo providing the desired methyl ester intermediate, 1-(((ferf-butoxy¡carbon¡l)amino)methyl)cyclopropanecarbox¡ methyl latate (0.400 g, 75% yield).

Step 2: Synthesis of ((1-(hydroxy¡methyl)cycloprop¡l)methyl)ferf-butyl carbamate

Methyl 1-(((ferf-butoxy¡carbon¡l)amino)methyl)cyclopropanecarboxylate (1 equiv.) was dissolved in THF and cooled to 0 °C. Lithium aluminum hydride (3 equiv.) was added slowly to the container, and the contents were stirred while allowed to warm to 23 °C over a period of 4 h. The reaction solution was then cooled to 0 °C and then water (x ml water/x g L¡AlH4 used), 15% sodium hydroxide solution (x ml water/x g L¡ AlH4 used) and water (3 x ml of water/x g of L¡AlH4 used) to the reaction sequentially. The reaction was filtered through celite and the filtrate was concentrated in vacuo. The residue was purified by silica gel chromatography providing the desired alcohol intermediate, ferf-butyl ((1-(hydroxymethyl)cyclopropyl)methyl)carbamate (0.41 g). , performance 88%).

Step 3: Synthesis of ((1-formylc¡cloprop¡l)methyl)ferf-butyl carbamate

To a solution of ((1-(hydroxy¡methyl)cycloprop¡l)methyl)ferf-butyl carbamate (1 equiv.) in dichloromethane at 25 °C was added PCC (1.15 equiv. ) in one part. The reaction was stirred for 2 h. Diethyl ether was added to the vessel, and the heterogeneous mixture was filtered through silica gel to provide the desired aldehyde intermediate, which was used without further purification.

Stage 4: Synthesis of (1-(1H-¡m¡dazol-2-yl)cycloprop¡l)methanam¡ne

A stirred solution of ferf-butyl ((1-formylcyclopropyl)methyl)carbamate (1 equiv.) in methanol was treated with ammonium hydroxide (10 equiv.) followed by oxalaldehyde (1.1 equiv. ). The contents were allowed to stir at 23 °C for 3 hours before removing the methanol in vacuo. The residue was then treated with TFA in dichloromethane (1:1 ratio) and stirred at 23 °C for 5 h. The mixture was diluted with brine and extracted with dichloromethane. The organic substances were dried over sodium sulfate and concentrated in vacuo to provide the desired imidazole intermediate, (1-(1H-ímídazol-2-yl)cyclopropíl)methanamíne (0.124 g, 100% yield), which was collected in the next reaction without further purification.

Stage 4: Synthesis of reference compound I-567

The title compound was prepared following general procedure B, except that (1-(1H-imidazole-2-¡l)cycloprop¡l)methanam¡ne was the amine reagent, and the contents were heated to 110 °C for 24 h as a solution in THF/water (10:1). The contents were cooled to 23 °C, and the organic solvents were removed in vacuo. The residue was purified by reverse phase HPLC to provide the desired product, reference compound I-567 (36 mg, 27% yield) as a white solid.

1H NMR (400 MHz, CDCla) 8 ppm 9.16 (br. s., 1 H), 8.53 (d, 1 H), 7.98 - 8.05 (m, 1 H), 7.51 (s, 1 H), 7.22 - 7.31 (m, 2 H), 6.97 - 7.09 (m, 2 H), 6.90 (s, 2 H), 6.81 (d , 2 H), 5.92 (s, 2 H), 4.05 (d, 2 H), 1.40 - 1.47 (m, 2 H), 1.32 - 1.39 (m, 2 H).

REFERENCE COMPOSITION I - 589

A solution of 5-fluoro-2-(1-(2-fluorobenzyl)-5-(isoxazol-3-yl)-1H--pyrazol-3-yl)pyrimidin-4-amine (intermediate product described in the previous patent : WO2012/3405 A1) (1 equiv.), 2-(methylsulfonyl)propanoic acid (3 equiv.), triethylamine (10 equiv.), and 2,4,6-tripropyl-1-2,4,6-trioxide ,3,5,2,4,6-trioxatriphosphinane (4 equiv.) in DMF was heated at 90 °C for 4 h. The reaction was cooled to 23 °C, then poured into a 1:1 mixture of ethyl acetate and water. The layers were separated and the aqueous layer was extracted with ethyl acetate (2 times). The organic substances were washed with water (3 times), dried over magnesium sulfate, filtered, and concentrated in vacuo. The resulting residue was purified by reverse phase HPLC to provide the desired product, reference compound I-589 (10 mg, 28% yield) as a white solid.

1H NMR (500 MHz, DMSO-da) 8 ppm 11.49 (s, 1 H), 9.11 (d, 1 H), 8.92 (d, 1 H), 7.61 (s, 1 H) ), 7.31 - 7.37 (m, 1 H), 7.27 (d, 1 H), 7.19 - 7.25 (m, 1 H), 7.12 (td, 1 H), 6.92 - 6.97 (m, 1 H), 5.92 (s, 2 H), 4.40 (d, 1 H), 3.07 (s, 3 H), 1.58 (d, 3H).

Reference compound I-608

The title compound was prepared in 4 steps:

Stage 1: Synthesis of ferf-butyl (2-hydrazinyl-2-oxoethyl)carbamate.

To a solution of methyl 2-((ferf-butoxycarbonyl)amino)acetate (1 equiv.) in ethanol was added hydrazine hydrate (15 equiv.), and the reaction was allowed to stir overnight. The solvent was removed in vacuo and the residue was triturated with hexane, filtered and dried under high vacuum to provide the desired acylhydrazine intermediate (2-hydrazinyl-2-oxoethyl)ferf-butyl carbamate, intermediate B (0 .89 g, yield 92%) as a white solid.

Step 2: Synthesis of ferf-butyl (2-oxo-2-(2-(2,2,2-trifluoroacetyl)hydrazinyl)ethyl)carbamate

To a solution of ferf-butyl (2-hydrazinyl-2-oxoethyl)carbamate (1 equiv.) in acetonitrile was added DIEA (1.1 equiv.). The contents were cooled to -45 °C, and 2,2,2-trifluoroacetic anhydride (1.1 equiv.) was added to the reaction. The resulting mixture was stirred while slowly heating to 23 °C. The solvent was removed in vacuo and the residue was partitioned between water and ethyl acetate. The layers were separated and the aqueous layer was extracted with ethyl acetate. The combined organic phases were washed with water and brine, dried with Na2SO4, filtered and concentrated in vacuo. The residue was purified by silica gel chromatography, using a gradient of 5-45% ethyl acetate in hexanes providing the desired intermediate, (2-oxo-2-(2-(2,2,2-trifluoroacetyl) Ferf-butyl hydrazinyl)ethyl)carbamate (0.73 g, 54% yield).

Step 3: Synthesis of (5-(trifluoromethyl)-1,3,4-oxadiazol-2-yl)methanamine

To a suspension of ferf-butyl (2-oxo-2-(2-(2,2,2-trifluoroacetyl)hydrazinyl)ethyl)carbamate (1 equiv.) in acetonitrile, DIEA (5.8 equiv.) was added and triphenylphosphine (4.1 equiv.), which was stirred for 5 min. Perchloroethane (2.3 equiv.) was then added to the reaction, and the mixture was stirred for 20 h at 23 °C. The solvent was removed in vacuo and the residue was partitioned between water and ethyl acetate. The layers were separated and the aqueous layer was extracted with ethyl acetate. The combined organic phases were washed with water and brine, dried with Na2SO4, filtered and concentrated in vacuo. The residue was purified by silica gel chromatography, using a gradient of 5-45% ethyl acetate in hexanes providing the N-Boc protected oxadiazole intermediate, ((5-(trifluoromethyl)--1,3, Ferf-butyl 4-oxadiazol-2-yl)methyl)carbamate (0.24 g, 35% yield). To a stirred solution of this N-Boc protected oxadiazole intermediate (1 equiv.) in dichloromethane was added TFA (8 equiv.), and the mixture was stirred at 23 °C for 4 h. The solvent was removed in vacuo providing the desired free amine intermediate, (5-(trifluoromethyl)-1,3,4-oxadiazol-2-yl)methanamine (as HCl salt, 0.15 g, 100% yield). ), which was used in the next step without further purification.

Step 4: Composite Synthesis Reference I - 608

To a stirred solution of (5-(trifluoromethyl)-1,3,4-oxadiazol-2-yl)methanamine (as the HCl salt, 2 equiv.) in dioxane cooled to 0 °C was added cesium carbonate (3 equiv.), and the mixture was stirred for 1 h. Intermediate 1 (1 equiv.) was added to the reaction, and the resulting mixture was stirred at 90 °C for 24 h. The reaction was cooled to 23 °C and diluted with ethyl acetate. The organic substances were washed with water and brine, concentrated in vacuo, and the resulting residue was purified by reverse phase HPLC to provide the desired product, the reference compound I-608 (2.5 mg, 5% yield) as a white solid.

1H NMR (500 MHz, CD3OD) 8 ppm 8.67 (d, 1 H), 8.15 (d, 1 H), 7.31 (s, 1 H), 7.17 (ddd, 1H), 6 .96-7.01 (m, 1 H), 6.90 - 6.95 (m, 1 H), 6.76 (d, 1 H), 6.70 - 6.74 (m, 1 H) , 5.85 (s, 2 H), 5.09 (s, 2 H).

Reference compound I-622

The title compound was prepared in 4 steps:

Step 1: Synthesis of (R)-tert-butyl (1-hydrazinyl-1-oxopropan-2-yl)carbamate

The title compound was prepared according to the procedure described in step 1 for the synthesis of the reference compound I-608, except for the use of (R)-methyl 2-((tert-butoxycarbonyl)amino)propanoate as the starting material. departure (97% performance).

Step 2: Synthesis of (R)-tert-butyl (1-oxo-1-(2-(2,2,2-trifluoroacetyl)hydrazinyl)propan-2-yl)carbamate. It was prepared according to the procedure described in step 2 for the synthesis of the reference compound I-608, except for the use of (R)-tert-butyl (1-hydrazinyl-1-oxopropan-2-yl)carbamate as material. starting point (yield 82%).

Step 3: Synthesis of (R)-1-(5-(trifluoromethyl)-1,3,4-oxadiazol-2-yl)ethanamine

It was prepared according to the procedure described in step 3 for the synthesis of the reference compound I-608, except for the use of (1-oxo-1-(2-(2,2,2-trifluoroacetyl)hydrazinyl)propan-2 -il)

-(R)-tert-butyl carbamate as starting material (yield 37%).

Stage 4: Synthesis of reference compound I-622

To a stirred solution of (R)-1-(5-(trifluoromethyl)-1,3,4-oxadiazol-2-yl)ethanamine, (R)-1-(5-(trifluoromethyl)-1,3,4 -oxadiazol-2-yl)ethanamine (2 equiv.) and intermediate 1 (1 equiv.) In DMF, cesium carbonate (3 equiv.) was added. The mixture was heated to 90 °C and stirred for 24 h. The contents were cooled to 23 °C and diluted with ethyl acetate. The mixture was washed with water and brine, concentrated in vacuo, and the resulting residue was purified by reverse phase HPLC to provide the desired product, reference compound I-622 (5 mg, 9% yield) as a white solid. 1

1H NMR (500 MHz, CD3OD) 8 ppm 8.66 - 8.69 (m, 1 H), 8.14 (d, 1 H), 7.27 (s, 1 H), 7.15-7, 21 (m, 1 H), 6.99 (dd, 1 H), 6.93 (t, 1 H), 6.76 (d, 1 H), 6.72 (t, 1 H), 5, 87 - 5.91 (m, 1 H), 5.85 (s, 2 H), 1.74 (d, 3 H).

Reference compound I-616

To a stirred solution of 2-(methylsulfonyl)acetamide (1 equiv.) in DMF was added cesium carbonate (3 equiv.) at 0 °C, and the mixture was stirred for 1 h. Intermediate 1 (1 equiv.) was added to the vessel, and the reaction was heated to 90 °C and stirred for 24 h. The contents were cooled to 23 °C and diluted with ethyl acetate. The mixture was washed with water and brine, concentrated in vacuo, and the resulting residue was purified by reverse phase HPLC to provide the desired product, reference compound I-616 (11 mg, 22% yield) as a white solid.

1 H R M N (500 M H z, C D 3O D) 5 p p m 8, 67 (d, 1 H), 8, 59 (d, 1 H), 7, 45 (s, 1 H), 7, 17 (d d d, 1 H), 6, 91 - 7, 02 (m, 2 H), 6, 77 - 6, 82 (m, 2 H), 5, 87 (s, 2 H), 4, 58 (b r. s., 2H), 3, 11(s, 3H).

Reference compound I-386

The title compound was prepared following general procedure C, except that 1H-pyrazole-3-carboxylic acid was the acid reagent, and the crude material was purified by silica gel chromatography using a gradient of 3-8% methanol in dichloromethane providing the desired compound, reference compound I-386 (20.2 mg, 40% yield) as a light tan solid. 1H-NMR (500 MHz, CDCh) 5 ppm 9.93 (s, 1 H), 8.77 (d, 1 H), 8.50 (s, 1 H), 8.33 (d, 1 H) , 7.51 (s, 1 H), 7.44 (d, 1 H), 7.19-7.25 (m, 1 H), 7.02-7.08 (m, 1 H), 6 .96-7.02 (m, 1 H), 6.94 (d, 1H), 6.83-6.87 (m, 1 H), 6.65 (s, 1 H), 6.02 ( s, 2H); 1 N-H proton was not observed.

Reference compound I-164

To a solution of intermediate 2 (1 equiv.) in dichloromethane was added trifluoroacetic anhydride (3 equiv.) followed by triethylamine (3 equiv.). The reaction was heated at 60 °C for 20 min, after which the reaction was concentrated in vacuo. The crude material was purified by silica gel chromatography with a gradient of 1-3% methanol in dichloromethane providing the desired compound, reference compound I-635 (16.4 mg, 32% yield) as a white solid.

1H-NMR (500 MHz, CDCh) 58.95 (br. s, 1 H), 8.86 (d, 1 H), 8.49 (d, 1 H), 8.10 (d, 1 H) , 7.49 (s, 1 H), 7.20-7.26 (m, 1 H), 7.03-7.07 (m, 1 H), 6.98-7.02 (m, 1 H), 6.82-6.87 (m, 1H), 6.62 (d, 1 H), 6.04 (s, 2 H).

Reference compound I-458

The title compound was prepared following general procedure C, except that 3-hydroxy-5-oxocyclohex-3-encarboxylic acid (1.3 equiv.) was used as the acid reagent and 2.5 equivalents of T3P. The crude material was purified by silica gel chromatography with a gradient of 3-10% methanol in dichloromethane providing the desired compound, reference compound I-458 (26.4 mg, 30% yield) as a white solid.

1H-NMR (500 MHz, CD3OD) 58.79 (m, 1 H), 8.68 (d, 1 H), 8.13 (d, 1 H), 7.53 (s, 1 H), 7 .24-7.33 (m, 1 H), 7.08-7.13 (m, 1 H), 7.01-7.08 (m, 1 H), 6.86-6.92 (m , 2 H, 2 isochronous displacements), 5.97 (s, 2 H), 2.66-2.75 (m, 2 H), 2.56 2.64 (m, 2 H); no 1 C-H proton was observed (isochronous with the solvent peak).

Reference compound I-459

The title compound was prepared following general procedure C, except that 5-oxocyclohex-2-carboxylic acid (1.2 equiv.) was used as the acid reagent and 2.5 equivalents of T3P. The crude material was purified by silica gel chromatography with a gradient of 3-10% methanol in dichloromethane, followed by a second purification by silica gel chromatography with a gradient of 7-12% (7:1 methanol/acetonitrile). in dichloromethane providing the desired compound, reference compound I-459 (12.6 mg, 15% yield) as a white solid.

1H-NMR (500 MHz, CD3OD) 58.79 (s, 1 H), 8.70 (d, 1 H), 8.13 (d, 1 H), 7.52 (s, 1 H), 7 .26-7.32 (m, 1 H), 7.08-7.13 (m, 1 H), 7.02-7.08 (m, 1 H), 6.87-6.93 (m , 2 H, 2 isochronous movements), 5.95 (s, 2H), 4.41-4.49 (m, 1 H), 2.52 2.60 (m, 1 H), 2.40-2 .50 (m, 1 H), 2.32-2.40 (m, 1 H), 2.20-2.30 (m, 1 H).

Reference compound I-464

The title compound was prepared following general procedure C, except that 5-oxypyrrolidine-3-carboxylic acid (1.2 equiv.) was used as the acid reagent and 2.5 equivalents of T3P. The crude material was purified by silica gel chromatography with a gradient of 3-10% methanol in dichloromethane providing the desired compound, reference compound I-464 (31.3 mg, 31% yield) as a white solid.1

1H-NMR (500 MHz, CD3OD) 58.79 (s, 1 H), 8.69 (d, 1 H), 8.13 (d, 1 H), 7.51 (s, 1 H), 7 .26-7.31 (m, 1 H), 7.07-7.13 (m, 1 H), 7.02-7.07 (m, 1 H), 6.86-6.93 (m , 2 H, 2 isochronous movements), 5.97 (s, 2 H), 3.67-3.76 (m, 1 H), 3.57 3.65 (m, 2 H), 2.60- 2.72 (m, 2H).

Reference compound I-461

The title compound was prepared following general procedure C, except that 1-(benzyloxy)cyclopropanecarboxylic acid (1 equiv.) was the acid reagent and 2.5 equivalents of T3P were used. The crude material was purified by silica gel chromatography with a gradient of 30-50% ethyl acetate/hexanes providing the desired compound, reference compound I-461 (14.2 mg, 19% yield) as a solid. tan color.

1 H - R M N ( 500 MHz , C D C l a ) 5 9 , 33 ( s , 1 H ) , 8 , 73 ( d , 1 H ) , 8 , 48 ( d , 1 H ) , 8 , 08 ( d , 1 H), 7, 42 (s, 1 H), 7, 34 - 7, 4 1 (m, 4 H), 7, 29 - 7, 3 2 (m, 1 H), 7, 18 - 7, 2 3 (m, 1 H), 7, 0 2 - 7, 06 (m, 1 H), 6, 9 7 - 7, 0 1 (m, 1 H), 6, 84 - 6, 88 (m, 1 H), 6, 61 (d, 1 H), 6, 03 (s, 2 H), 4, 68 (s, 2 H), 1, 4 5 - 1, 5 1 (m, 2 H), 1 , 3 2 - 1 , 3 7 ( m , 2 H ) .

Reference compound I-469

The title compound was prepared following general procedure C, except that 2-(thiazol-2-yl)acetic acid was the acid reagent. The crude material was purified by silica gel chromatography with a gradient of 3-8% methanol in dichloromethane, followed by a second purification by reverse phase HPLC with a gradient of 10-95% acetonitrile in water providing the desired compound, the Reference compound I-469 (4.3 mg, 6% yield) as a tan solid.

1H-NMR (500 MHz, CD3OD) 88.81 (s, 1 H), 8.69 (d, 1 H), 8.20 (d, 1 H), 7.81 (d, 1 H), 7 .61 (d, 1 H), 7.57 (s, 1 H), 7.25 7.33 (m, 1 H), 7.08-7.13 (m, 1 H), 7.02- 7.07 (m, 1 H), 6.87-6.96 (m, 2 H, 2 isochronous movements), 5.99 (s, 2 H), 3.30 (s, 2 H).

Reference compound I-465

To a solution of 5-fluoro-2-(1-(2-fluorobenzyl)-5-(isoxazol-3-yl)-1H-pyrazol-3-yl)--pyrimidin-4-amine (intermediate product described in WO2012 /3405 A1; 1 equiv.) in dichloromethane, trifluoroacetic anhydride (3 equiv.) was added followed by triethylamine (3 equiv.). The reaction was heated at 60 °C for 20 min, after which the reaction was concentrated in vacuo. The crude material was purified by silica gel chromatography with a gradient of 1-3% methanol in dichloromethane providing the desired compound, reference compound I-465 (26.8 mg, 28% yield) as a white solid.

1H-NMR (500 MHz, CDCh) 88.77 (s, 1 H), 8.56 (br. s, 1 H), 8.40 (s, 1 H), 7.44 (s, 1 H) , 7.19-7.25 (m, 1 H), 7.02-7.08 (m, 1 H), 6.96-7.02 (m, 1 H), 6.81-6.88 (m, 1 H), 6.62 (d, 1 H), 6.02 (s, 2H).

Reference compound I-470

To a mixture of reference compound I-38 (1 equiv.), 1-hydroxycyclopropanecarboxylic acid (1.1 equiv.) and 4-dimethylaminopyridine (0.1 equiv.) in dichloromethane was added triethylamine (3 equiv.) followed by 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (1.1 equiv.). The reaction was stirred at room temperature for 12 h, after which the reaction was diluted with water and 1N hydrochloric acid solution and extracted with dichloromethane. The combined organic layers were dried over sodium sulfate, filtered, and concentrated in vacuo. The crude material was purified by reverse phase HPLC using a gradient of 10-95% acetonitrile in water providing the desired compound, reference compound I-470 (1.3 mg, 4% yield) as a white solid.

1H-NMR (500 MHz, CDCla) 88.50 (d, 1 H), 8.26 (s, 1 H), 7.33 (s, 1 H), 7.19-7.26 (m, 1 H), 6.97-7.07 (m, 3 H), 6.67 (m, 1 H), 5.93 (s, 2 H), 5.31 (m, 1 H), 4.91 -5.04 (m, 2H), 4.42-4.75 (m, 2H), 4.16-4.32 (m, 1H), 1.28-1.43 (m, 2H) ), 0.79-0.92 (m, 2 H); 1 exchangeable proton was not observed.

Reference compound I-471

The title compound was prepared following general procedure C, except that 3,3,3-trifluoropropanoic acid was the acid reagent, 2.5 equivalents of T3P were used, and the reaction was stirred at 23 °C for 24 h. The crude material was purified by silica gel chromatography using a gradient of 3-10% methanol in dichloromethane providing the desired compound, reference compound I-471 (79.3 mg, 85% yield) as a tan solid. .

1H-NMR (500 MHz, CDCh) 88.78 (d, 1 H), 8.48 (d, 1 H), 8.47 (br. s, 1 H), 8.09 (d, 1 H) , 8.03 (s, 1 H), 7.47 (s, 1 H), 7.19-7.24 (m, 1 H), 7.02-7.09 (m, 1 H), 6 .96-7.01 (m, 1 H), 6.81-6.86 (m, 1 H), 6.61 (d, 1 H), 6.03 (s, 1 H), 3.29 (q, 2H).

Reference compound I-472

To a solution of intermediate 2 (1 equiv.) in dichloromethane was added methylsulfonylmethylsulfonyl chloride (1.08 equiv.) followed by 1,8-diazabicyclo[5.4.0]undec-7-ene (1 equiv.). The reaction was heated to 60 °C for 1 h, after which the reaction was diluted with water and 1 N hydrochloric acid solution and extracted with dichloromethane. The combined organic layers were dried over sodium sulfate, filtered, and concentrated in vacuo. The crude material was purified by silica gel chromatography with a gradient of 3-8% methanol in dichloromethane providing the desired compound, reference compound I-472 (39.6 mg, 37% yield) as a white solid.1

1H-NMR (500 MHz, CDCh) 88.55 (d, 1 H), 8.26 (br. s, 1 H), 7.36 (s, 1 H), 7.26-7.30 (m , 1 H), 7.07-7.16 (m, 3 H), 6.84-6.91 (m, 1 H), 6.62-6.67 (m, 1 H), 5.95 (s, 2 H), 4.60 (s, 2 H), 3.17 (s, 3 H); 1 N-H proton was not observed.

Reference compound I-486

The title compound was prepared following general procedure C, except that 4-sulfamoylbutanoic acid was the acid reagent, and 2.5 equivalents of T3P were used. The crude material was purified by silica gel chromatography with a gradient of 3-10% methanol in dichloromethane providing the desired compound, reference compound I-486 (14.7 mg, 15% yield) as a white solid.

1 H - R M N ( 500 M H z , C D 3O D ) 58 .78 (s, 1H ) , 8 , 66 ( d , 1 H ) , 8 , 12 ( d , 1 H ) , 7 .52 ( s , 1 H ) , 7 , 25 - 7 , 3 2 ( m , 1 H ) , 7 , 0 7 - 7 , 13 ( m , 1 H ) , 7 , 0 2 - 7 , 0 7 ( m , 1 H ) , 6 , 86 - 6, 91 (m, 2 H, 2 isochronous displacements), 5, 97 (s, 2 H), 3, 19 (t, 2 H), 2, 71 (t, 2 H), 2, 21 (m, 2H).

Reference compound I-496

To 0 °C suspension of intermediate 2 (1 equiv.) in dichloromethane was added trimethylaluminum (2 M solution in toluene, 0.45 equiv.). The reaction was heated to 23 °C after which α,α-dimethyl-Y-butyrolactone (1.1 equiv.) was added. The reaction was heated to 80 °C for 16 h, cooled to 23 °C, then diluted with saturated ammonium chloride solution, extracted with ethyl acetate, and washed with 1N hydrochloric acid solution. The combined organic layers were dried over sodium sulfate, filtered, and concentrated in vacuo. The crude material was purified by reverse phase HPLC using a gradient of 5-75% of an acetonitrile in water providing the desired compound, reference compound I-496 (7.7 mg, 25% yield) as a white solid.

1H-NMR (500 MHz, CDCh) 58.71 (d, 1 H), 8.47 (d, 1 H), 8.34 (d, 1 H), 7.41 (s, 1 H), 7 .17-7.24 (m, 1 H), 7.01-7.07 (m, 1 H), 6.95-6.99 (m, 1 H), 6.85-6.90 (m , 1 H), 6.60 (d, 1 H), 6.00 (s, 2 H), 4.15 (t, 2 H), 2.04 (t, 2 H), 1.29 (s , 6H).

Reference compound I-501

To a mixture of intermediate 1 (1 equiv.) and 5-(trifluoromethyl)pyrrolidin-2-one (1.2 equiv.) in 1,4-dioxane was added cesium carbonate (1.5 equiv.). The reaction was heated at 100 °C for 16 h, after which the reaction was diluted with water, extracted with dichloromethane, and washed with saturated sodium bicarbonate solution. The combined organic layers were dried over sodium sulfate, filtered, and concentrated in vacuo. The crude material was purified by reverse phase HPLC using a gradient of 10-95% acetonitrile in water providing the desired compound, reference compound I-501 (13.4 mg, 13% yield) as an off-white solid. 1H-NMR (500 MHz, CDCla) 58.76 (d, 1 H), 8.49 (d, 1 H), 7.38 (s, 1 H), 7.21-7.26 (m, 1 H), 7.03-7.07 (m, 1 H), 6.98-7.02 (m, 1 H), 6.86-6.92 (m, 1 H), 6.61 (d , 1 H), 6.01 (s, 2 H), 5.34-5.39 (m, 1 H), 2.88-2.99 (m, 1 H), 2.58-2.70 (m, 2 H), 2.40-2.46 (m, 1 H).

Reference compound I-503

To a mixture of intermediate 1 (1 equiv.) and 1,1-isothiazolidine dioxide (1.2 equiv.) in 1,4-dioxane was added cesium carbonate (1.5 equiv.). The reaction was heated at 100 °C for 16 h, after which the reaction was diluted with water, and extracted with ethyl acetate. The combined organic layers were dried over sodium sulfate, filtered, and concentrated in vacuo. The crude material was purified by silica gel chromatography with a gradient of 3-10% methanol in dichloromethane providing the desired compound, reference compound I-503 (71.3 mg, 73% yield) as an off-white solid.

1H-NMR (500 MHz, CDCh) 58.57 (d, 1 H), 8.47 (d, 1 H), 7.36 (s, 1 H), 7.20-7.25 (m, 1 H), 7.03-7.07 (m, 1 H), 6.96 7.01 (m, 1 H), 6.84-6.88 (m, 1 H), 6.61 (m, 1 H), 5.99 (s, 2 H), 4.27 (t, 2 H), 3.44 (t, 2 H), 2.66 (t, 2 H).

Reference compound I-506

To a mixture of intermediate 1 (1 equiv.) and piperidin-2-one (1.2 equiv.) in 1,4-dioxane was added cesium carbonate (1.5 equiv.). The reaction was heated at 100 °C for 16 h, after which the reaction was diluted with water, extracted with dichloromethane, and washed with 1N sodium hydroxide solution. The combined organic layers were dried over sodium sulfate, filtered, and concentrated in vacuo. The crude material was purified by reverse phase HPLC using a gradient of 5-95% of an acetonitrile in water providing the desired compound, the compound reference I-506 (4.9 mg, 5% yield) as a white solid.

1H-NMR (500 MHz, CDCh) 58.64 (d, 1 H), 8.48 (d, 1 H), 7.38 (s, 1 H), 7.19-7.23 (m, 1 H), 7.03-7.07 (m, 1 H), 6.97 7.01 (m, 1 H), 6.85-6.88 (m, 1 H), 6.60 (d, 1 H), 6.00 (s, 2 H), 4.00 (t, 2 H), 2.65 (t, 2 H), 1.98-2.07 (m, 4 H).

Reference compound I-512

To a mixture of intermediate 1 (1 equiv.) and 5,5-dimethylpyrrolidin-2-one (1.2 equiv.) in 1,4-dioxane was added cesium carbonate (1.5 equiv.). The reaction was heated at 100 °C for 16 h, after which the reaction was diluted with water, extracted with dichloromethane, and washed with saturated sodium bicarbonate solution. The combined organic layers were dried over sodium sulfate, filtered, and concentrated in vacuo. The crude material was purified by reverse phase HPLC using a gradient of 10-95% acetonitrile in water providing the desired compound, reference compound I-512 (0.6 mg, 1% yield) as an off-white solid.

1H-NMR (500 MHz, CDCh) 58.69 (d, 1 H), 8.49 (d, 1 H), 7.28 (s, 1 H), 7.20-7.25 (m, 1 H), 7.01-7.06 (m, 1 H), 6.98 7.01 (m, 1 H), 6.89-6.95 (m, 1 H), 6.60 (d, 1 H), 5.97 (s, 2 H), 2.67 (t, 2 H), 2.14 (t, 2 H), 1.63 (s, 6 H).

Reference compound I-526 and reference compound I-527

A mixture of the intermediate product 1 (1 eq u iv .) and a mixture of 3 - m e t i l - 3 - (m e t i l su l f o n i l ) p i r r o l i d i n - 2 - o n a y 4 - h i d r o x i - 2 - m e t i l - 2 - (m e t i l su l f o n i l ) - - b u t a n a m i d a (com b in a d a, 1 eq u iv ) in 1, 4 - d i o x a n o b e a d i o c a r b o n a t o d e c e s i o (1.5 equiv.). The reaction was heated at 100 °C for 16 h, after which the reaction was diluted with water, and extracted with ethyl acetate. The combined organic layers were dried over sodium sulfate, filtered, and concentrated in vacuo. The crude material was purified by reverse phase HPLC using a gradient of 5-95% acetonitrile in water providing the desired compounds, reference compound I-526 (0.5 mg, 2% yield) and reference compound I- 527 (1.3 mg, 5% yield) as a white solid.

1H-NMR for Reference Compound I-526 (500 MHz, CDCh) 88.73 (d, 1 H), 8.49 (d, 1 H), 7.37 (s, 1 H), 7.20 7.25 (m, 1 H), 7.02-7.06 (m, 1 H), 6.96-7.01 (m, 1 H), 6.84-6.89 (m, 1 H) ), 6.61 (d, 1 H), 6.01 (s, 2 H), 4.28-4.33 (m, 1 H), 4.18-4.23 (m, 1 H), 3.20-3.25 (m, 1 H), 3.12 (s, 3 H), 2.29-2.35 (m, 1 H), 2.87 (s, 3 H).

1H-NMR for Reference Compound I-527 (500 MHz, CDCla) 88.50 (d, 1 H), 8.44 (d, 1 H), 7.38 (s, 1 H), 7.20 7.26 (m, 1 H), 7.06-7.11 (m, 1 H), 7.02-7.06 (m, 1 H), 6.95-7.03 (m, 1 H) ), 6.65 (d, 1 H), 6.00 (d, 1 H), 5.98 (d, 1 H), 5.91 (br. s, 1 H), 4.82-4, 86 (m, 1 H), 4.74-4.78 (m, 1 H), 3.03 (s, 3 H), 2.86-2.90 (m, 1 H), 2.48- 2.52 (m, 1 H), 1.76 (s, 3 H).

Reference compound I-533

To a mixture of intermediate 1 (1 equiv.) and pyrrolidin-2,5-one (1.3 equiv.) in 1,4-dioxane was added cesium carbonate (1.5 equiv.). The reaction was heated at 100 °C for 16 h, after which the reaction was diluted with water, extracted with ethyl acetate, and washed with 1N sodium hydroxide solution. The combined organic layers were dried over sodium sulfate, filtered, and concentrated in vacuo. The crude material was purified by reverse phase HPLC using a gradient of 5-95% acetonitrile in water providing the desired compound, reference compound I-533 (3.8 mg, 5% yield) as a white solid.

1H-NMR (500 MHz, CDCh) 88.86 (d, 1 H), 8.47 (d, 1 H), 7.42 (s, 1 H), 7.20-7.24 (m, 1 H), 7.01-7.06 (m, 1 H), 6.96 7.00 (m, 1 H), 6.81-6.85 (m, 1 H), 6.59 (d, 1H), 6.02 (s, 2H), 3.02 (s, 4H).

Reference compound I-534

To a mixture of intermediate 1 (1 equiv.) and 5-oxypyrrolidine-2-carboxamide (1.2 equiv.) in 1,4-dioxane was added cesium carbonate (1.5 equiv.). The reaction was heated at 100 °C for 24 h, after which the reaction was diluted with water and extracted with ethyl acetate. The combined organic layers were dried over sodium sulfate, filtered, and concentrated in vacuo. The crude material was purified by reverse phase HPLC using a gradient of 5-75% acetonitrile in water providing the desired compound, reference compound I-534 (0.6 mg, 1% yield) as a white solid.

1H-NMR (500 MHz, CDCh) 88.62 (d, 1 H), 8.50 (d, 1 H), 7.31 (s, 1 H), 6.99-7.07 (m, 3 H), 6.61 (d, 1 H), 6.01 (d, 1 H), 5.85 (d, 2 H), 5.29 (s, 2 H), 4.92-4.96 (m, 1 H), 2.87-2.93 (m, 1 H), 2.58-2.63 (m, 1 H), 2.43-2.55 (m, 2 H).

Reference compound I-590

To a mixture of intermediate 1 (1 equiv.) and 5-oxypyrrolidine-3-carboxamide (1.3 equiv.) in 1,4-dioxane was added cesium carbonate (1.5 equiv.). The reaction was heated at 100 °C for 16 h, after which the reaction was diluted with water, extracted with ethyl acetate, and washed with 1N hydrochloric acid solution. The combined organic layers were dried over sodium sulfate, filtered, and concentrated in vacuo. The crude material was purified by silica gel chromatography with a gradient of 5-12% methanol in dichloromethane providing the desired compound, reference compound I-590 (3.2 mg, 3% yield) as a tan solid. .

1H-NMR (500 MHz, CD3OD) 88.76 (d, 1 H), 8.74 (d, 1 H), 7.53 (s, 1 H), 7.25-7.29 (m, 1 H), 7.07-7.11 (m, 1 H), 7.02 7.07 (m, 1 H), 6.91 (d, 1 H), 6.83-6.87 (m, 1 H), 5.97 (s, 2 H), 4.26-4.37 (m, 2 H), 3.45-3.49 (m, 1 H), 2.84-2.94 ( m, 2H).

Reference compound I-691

To a mixture of intermediate 1 (1 equiv.) and ethyl 3-methyl-2-oxopyrrolidine-3-carboxylate (1.2 equiv.) in 1,4-dioxane was added cesium carbonate (1.5 equiv. ). The reaction was heated to 75 °C for 16 h, after which the reaction was diluted with saturated ammonium chloride solution and extracted with dichloromethane. The combined organic layers were dried over sodium sulfate, filtered, and concentrated in vacuo. The crude material was purified by silica gel chromatography with a gradient of 3-7% methanol in dichloromethane providing the desired compound, reference compound I-691 (377 mg, 92% yield) as a white solid.

1H-NMR (500 MHz, CDCh) 88.68 (d, 1 H), 8.48 (d, 1 H), 7.37 (s, 1 H), 7.19-7.25 (m, 1 H), 7.01-7.06 (m, 1 H), 6.96 7.01 (m, 1 H), 6.86-6.90 (m, 1 H), 6.60 (d, 1 H), 6.00 (s, 2 H), 4.26 (q, 2 H), 4.15-4.21 (m, 2 H), 2.75-2.80 (m, 1 H ), 2.17-2.23 (m, 1 H), 1.59 (s, 3 H), 1.30 (t, 3H).

Reference compound I-604

Even a solution of the reference compound I - 591 (1 equ iv.) was added u o s a 1 M d e s o d i o h y d r oxide (2 eq u iv .) . The reaction was stirred at room temperature for 2 h, after which the reaction was concentrated to ~50% of its volume, acidified by adding a 1 M aqueous solution of hydrochloric acid and extracted with acetate. ethyl to . The combined organic layers were dried over sodium sulfate, filtered, and concentrated in vacuo to provide the desired compound, reference compound I-604 (154.6 mg, 95% yield) as a white solid.

1H-NMR (500 MHz, CDCh) 88.77-8.80 (m, 2 H, 2 overlapping movements), 7.57 (s, 1 H), 7.28-7.32 (m, 1 H) , 7.09-7.13 (m, 1 H), 7.04-7.09 (m, 1 H), 6.94 (d, 1 H), 6.86-6.90 (m, 1 H), 6.00 (s, 2 H), 4.26-4.31 (m, 1 H), 4.16-4.20 (m, 1 H), 2.75-2.79 (m , 1 H), 2.27-2.31 (m, 1 H), 1.54 (s, 3 H).

Reference compound I-605

To a -78 °C solution of reference compound I-604 (1 equiv.) in dichloromethane was added oxalyl chloride (2 M solution in dichloromethane, 2.5 equiv.). The reaction was stirred at −78 °C for 30 min, then warmed to 0 °C and stirred at that temperature for 1 h. The reaction was then concentrated in vacuo, reconstituted in dichloromethane and cooled to -78°C. Cyclopropylamine (5 equiv.) was added to this solution, after which the reaction was allowed to warm to room temperature. After 20 min, the reaction was concentrated in vacuo. The crude material was purified by silica gel chromatography with a gradient of 1-8% methanol in dichloromethane providing the desired compound, reference compound I-604 (14.5 mg, 23% yield) as a white solid.

1H-NMR (500 MHz, CDCla) 88.70 (d, 1 H), 8.49 (d, 1 H), 7.36 (s, 1 H), 7.20-7.25 (m, 1 H), 7.01-7.08 (m, 1 H), 6.97 7.01 (m, 1 H), 6.86-6.90 (m, 1 H), 6.60 (d, 1 H), 6.00 (s, 2 H), 4.06-4.09 (m, 2 H), 3.00-3.06 (m, 1 H), 2.75-2.80 ( m, 1 H), 2.14-2.29 (m, 1 H), 1.58 (s, 3 H), 0.78-0.82 (m, 2 H), 0.51-0, 54 (m, 2H); 1 N-H proton was not observed.

Reference compound I-606

To a -78°C solution of reference compound I-604 (1 equiv.) in dichloromethane was added a 2 M dichloromethane solution of oxalyl chloride (2.5 equiv.). The reaction was stirred at -78 °C for 30 minutes, then warmed to 0 °C, and stirred at that temperature for 1 h. The reaction was then concentrated in vacuo, reconstituted in dichloromethane, and cooled to -78°C. Ammonium hydroxide solution (50 equiv.) was added to this solution, after which the reaction was allowed to warm to room temperature. After 20 minutes, the reaction was diluted in water and extracted with ethyl acetate. The combined organic layers were dried over sodium sulfate, filtered, and concentrated in vacuo to provide the desired compound, reference compound I-619 (43.3 mg, 75% yield) as an off-white solid.

1H-NMR (500 MHz, CDCh) 88.71 (d, 1 H), 8.49 (d, 1 H), 7.37 (s, 1 H), 7.20-7.25 (m, 1 H), 7.03-7.07 (m, 1 H), 6.97 7.01 (m, 1 H), 6.86-6.91 (m, 1 H), 6.60 (d, 1 H), 6.00 (s, 2 H), 4.07-4.13 (m, 2 H), 2.97-3.03 (m, 1 H), 2.17-2.22 ( m, 1H), 1.65 (s, 3H); 2 N-H protons were not observed.

Reference compound I-612

A suspension of intermediate 1 (1 equiv) and potassium ((2-carboxylateethyl)sulfonyl)amide (1.15 equiv) in DMSO was stirred at room temperature for 72 h. The reaction was diluted in water, washed with dichloromethane, acidified by adding 1 M hydrochloric acid solution and extracted with ethyl acetate. The combined organic layers were dried over sodium sulfate, filtered, and concentrated in vacuo to provide the acidic intermediate. To a suspension of this acidic intermediate in dichloromethane was added triethylamine (3 equiv.), followed by oxalyl chloride (2 M solution in dichloromethane, 2 equiv.). After 15 min, the reaction was concentrated in vacuo. The crude material was purified by reverse phase HPLC using a gradient of 5-75% acetonitrile in water providing the desired compound, reference compound I-612 (5.8 mg, 12% yield) as a white solid.

1H-NMR (500 MHz, CDCh) 88.86 (d, 1 H), 8.47 (d, 1 H), 7.43 (s, 1 H), 7.19-7.24 (m, 1 H), 7.01-7.06 (m, 1 H), 6.96 7.01 (m, 1 H), 6.83-6.88 (m, 1 H), 6.63 (d, 1 H), 6.02 (s, 2 H), 3.89 (t, 2 H), 3.35 (t, 2 H).

Reference compound I-615

To a mixture of intermediate 1 (1 equiv.) and 3-dimethylpyrrolidin-2-one (1.2 equiv.) in 1,4-dioxane was added cesium carbonate (1.5 equiv.). The reaction was heated to 70 °C for 12 h, after which the reaction was diluted with water and extracted with ethyl acetate. The combined organic layers were dried over sodium sulfate, filtered, and concentrated in vacuo to provide the desired compound, reference compound I-615 (59.7 mg, 48% yield) as a white solid.

1H-NMR (500 MHz, CDCh) 88.48 (d, 1 H), 8.46 (d, 1 H), 7.32 (s, 1 H), 7.19-7.24 (m, 1 H), 7.02-7.07 (m, 1 H), 6.98 7.02 (m, 1 H), 6.89-6.94 (m, 1 H), 6.60 (d, 1 H), 5.97 (s, 2 H), 5.92-5.96 (m, 1 H), 3.60-3.65 (m, 1 H), 3.47-3.52 ( m, 1 H), 2.82-2.86 (m, 1 H), 2.33-2.41 (m, 1 H); 1 O-H proton was not observed.

Reference compound I-628

A h y d r a t i o ch l o r u c i o n o f 1 - (2 - f l u o ro b e n c i l ) - 5 - ( i s o x a z o l - 3 - i l ) - 1 H - p i r a z o l - 3 - c a r b o x i m i d a m i d e ( g e n e r a d In stage 3 of the general management procedure A, using 1 - (is o x a z o l - 3 - i l ) et a n o na in step 1 and 2 - f l u o ro b e n z i l h i d r a z i n a in step 2, 1 equiv.), methyl 4-oxotetrahydrothiophen-3-carboxylate (3 equiv.), and 1,8-diazabicyclo[5.4.0]undec-7- ene (1 equiv.) in pyridine was heated at 80 °C for 12 h. The reaction was concentrated in vacuo, suspended in methanol, concentrated in vacuo, and resuspended in methanol. The precipitate was filtered and dried to provide the desired cyclic sulfide intermediate (190 mg, 45% yield) as a light tan solid. To a solution of this sulfide intermediate (1 equiv.) in dichloromethane was added peracetic acid (2.3 equiv.). After 30 min, the reaction was concentrated in vacuo, suspended in water and filtered to provide the desired compound, reference compound I-628 (148.8 mg, 73% yield) as an off-white solid.

1H-NMR (500 MHz, CDCla) 810.2 (br. s, 1 H), 8.56 (s, 1 H), 7.31-7.34 (m, 1 H), 7.30 (s , 1 H), 7.07-7.12 (m, 3 H), 6.64 (m, 1 H), 5.93 (s, 2 H), 4.36 (s, 2 H), 4 .35 (s, 2 H).

Reference compound I-632

A suspension of reference compound I-628 (1 equiv.) in phosphorus oxychloride (62 equiv.) was heated at 90 °C for 2 h, after which the reaction mixture was concentrated in vacuo providing the chloropyrimidine intermediate. desired (155 mg, 100% yield) as a tan solid. To a suspension of this intermediate product (1 equiv.) in dioxane was added a solution of ammonium hydroxide (440 equiv.). The reaction was stirred at 23 °C for 15 h, then heated to 60 °C for 1 h, after which the mixture was diluted in water and extracted with ethyl acetate. The combined organic layers were dried over sodium sulfate, filtered, and concentrated in vacuo to provide the desired compound, reference compound I-632 (44.5 mg, 60% yield) as a tan solid. 1H-NMR (500 MHz, DMSO-de) 89.09 (d, 1 H), 7.51 (s, 1 H), 7.31-7.35 (m, 1 H), 7.27 (d , 1 H), 7.21-7.24 (m, 1 H), 7.09-7.13 (m, 1 H), 6.83-6.87 (m, 1 H), 5.90 (s, 2 H), 4.49 (s, 2 H), 4.31 (s, 2 H).

Reference compounds I-497 and I-524

A solution of intermediate 2 (1 equiv.), triethylamine (3.5 equiv.), DMAP (0.1 equiv.), and 2-chloro-2-oxethylacetate (2.2 equiv.) in dichloromethane was heated to 60 °C for 26 h. The solvent was removed in vacuo and the crude material was purified by reverse phase HPLC providing the desired compound, reference compound I-497 (30 mg, 23% yield) as a white solid, and a secondary product, the compound reference I-524 (4.5 mg, 4% yield) as a white solid.

1H-NMR for Reference Compound I-497 (500 MHz, DMSO-d6) 8 ppm 11.37 (m, 1 H), 9.11 (d, 1 H), 8.75 (d, 1 H) , 7.94 (m, 1 H), 7.66 (s, 1 H), 7.35 (m, 1 H), 7.27 (d, 2 H), 7.11 (m, 1 H) , 6.89 (m, 1 H), 5.93 (s, 2 H), 4.77 (s, 2 H), 2.12 (s, 3 H).

1H-NMR for Reference Compound I-524 (500 MHz, DMSO-d6) 8 ppm 11.13 (m, 1 H), 9.09 (m, 1 H), 8.72 (m, 1 H) , 8.00 (m, 1 H), 7.65 (s, 1 H), 7.35 (m, 1 H), 7.26 (s, 2 H), 7.12 (m, 1 H) , 6.88 (m, 1 H), 5.93 (s, 2 H), 2.15 (s, 3 H).

Reference compound I-499

To a suspension of the reference compound I-497 (1 equiv.) in methanol was added a solution of potassium carbonate (0.5 equiv.) in water. After stirring for 1 h at 23 °C, an additional 0.5 equivalent of potassium carbonate in water was added to the vessel, along with THF (volume equal to the starting volume of methanol). The reaction was left stirring for an additional 1 h at 23 °C. The solvent was removed in vacuo and the resulting crude material was purified by silica gel chromatography using a gradient of 0-5% methanol in dichloromethane providing the desired compound, reference compound I-499 (10.5 mg, yield 17 %) as a white solid.

1H-NMR (500 MHz, DMSO-d6) 8 ppm 10.43 (m, 1 H), 9.11 (m, 1 H), 8.75 (m, 1 H), 8.01 (m, 1 H), 7.68 (s, 1 H), 7.34 (m, 1 H), 7.24 (m, 2 H), 7.12 (m, 1 H), 6.89 (m, 1 H), 5.93 (s, 2 H), 5.61 (m, 1 H), 4.11 (m, 2 H).

Reference compound I-525

The title compound was prepared following general procedure C in the library format, except that 4-(benzyloxy)tetrahydro-2H-pyran-4-carboxylic acid was the acidic reagent, and the crude material was purified by phase HPLC. reverse providing the desired compound, reference compound I-525 (9 mg, 27% yield) as a white solid.

1H-NMR (500 MHz, DMSO-d6) 8 ppm 9.39 (m, 1 H), 8.91 (m, 1 H), 8.56 (s, 1 H), 8.36 (m, 1 H), 7.53 (m, 1 H), 7.38 (m, 5 H), 7.26 (m, 1H), 7.07 (s, 3 H), 6.65 (s, 1 H) ), 6.02 (s, 2 H), 4.51 (s, 2 H), 3.96 (m, 2 H), 3.87 (m, 2 H), 2.28 (m, 2 H ), 1.99 (m, 2H).

Reference compound I-528

The composition of the title was prepared following the general management procedure in the library format, except for acid 2 - method x The acidic reagent was the acidic reagent, and the raw material was purified by reverse-phase HPL C by providing the component. e a d o, the reference compound I-528 (7 mg, yield 58%) as a white solid.

1 H - R M N ( 500 MHz , D M S O - d a ) 5 p p m 9 , 18 ( m , 1 H ) , 8 , 75 ( m , 1 H ) , 8 , 47 ( m , 1 H ) , 8 , 18 ( m , 1 H ) , 7 , 50 ( m , 1 H ) , 7 , 21 ( m ,

1H), 7, 05 (m, 1H), 6, 97 (m, 1H), 6, 83 (m, 1H), 6, 61 (m, 1H), 6, 05 (m, 2 H), 4, 08 (m, 2 H), 3, 52 (s, 3 H).

Reference compound I-532

The title compound was prepared following general procedure C in the library format, except that oxazole-4-carboxylic acid was the acid reagent, and the crude material was purified by reverse phase HPLC providing the desired compound, the compound reference I-532 (3.8 mg, 15% yield) as a white solid.

1H-NMR (500 MHz, DMSO-d6) 5 ppm 10.39 (m, 1 H), 9.09 (m, 2 H), 8.82 (m, 1 H), 8.65 (m, 1 H), 8.11 (m, 1H), 7.73

(m, 1 H), 7.29 (m, 3 H), 7.13 (m, 1 H), 6.88 (m, 1 H), 5.94 (m, 2 H).

Reference compound I-547

The title compound was prepared following general procedure C in the library format, except that the acid

3-Methoxypropanoic acid was the acid reagent, and the crude material was purified by reverse phase HPLC to provide the desired compound, reference compound I-547 (4.9 mg, 20% yield) as a white solid.

1H-NMR (500 MHz, DMSO-d6) 5 ppm 11.09 (m, 1 H), 9.09 (m, 1 H), 8.72 (m, 1 H), 8.02 (m, 1 H), 7.64 (s, 1 H), 7.34 (m, 1 H), 7.23 (m, 2 H), 7.12 (m, 6.89 (m, 1 H), 5 .92 (s, 2 H), 3.61 (t, 2 H), 3.23 (s, 3 H), 2.70 (t, 2 H).

Reference compound I-548

The title compound was prepared following general procedure C in the library format, except that tosylalanine was the acidic reagent, and the crude material was purified by reverse phase HPLC to provide the desired compound, reference compound I-548 ( 3.1 mg, yield 9%) as a white solid.

Reference compound I-549

The title compound was prepared following general procedure C in the library format, except that thiazole-4-carboxylic acid was the acid reagent, and the crude material was purified by reverse phase HPLC providing the desired compound, the compound reference I-549 (3.7 mg, 14% yield) as a white solid.

1H-NMR (500 MHz, DMSO-d6) 5 ppm 10.20 (m, 1 H), 9.31 (m, 1 H), 9.09 (m, 1 H), 8.84 (m, 1 H), 8.72 (m, 1 H), 8.14 (m, 1 H), 7.73 (m, 1 H), 7.29 (m, 3 H), 7.13 (m, 1 H), 6.89 (m, 1 H), 5.94 (m, 2 H).

Reference compound I-550

The title compound was prepared following general procedure C in the library format, except that the acid

1H-pyrrole-2-carboxylic acid was the acid reagent, and the crude material was purified by reverse phase HPLC to provide the desired compound, reference compound I-550 (3.4 mg, 13% yield) as a white solid.

1H-NMR (500 MHz, DMSO-d6) 5 ppm 11.84 (m, 1 H), 10.84 (m, 1 H), 9.09 (m, 1 H), 8.73 (m, 1 H), 8.16 (m, 1H), 7.68

(s, 1 H), 7.36 (m, 2 H), 7.24 (m, 2 H), 7.13 (m, 1 H), 7.06 (m, 1 H), 6.88 (m, 1 H), 6.19 (m, 1 H), 5.94 (s, 2 H).

Reference compound I-551

The title compound was prepared following general procedure C in the library format, except that the acid

1-Cyanocyclopropane-1-carboxylic acid was the acid reagent, and the crude material was purified by reverse-phase HPLC to provide the desired compound, reference compound I-551 (3.3 mg, 13% yield) as a white solid.

1H-NMR (500 MHz, DMSO-d6) 5 ppm 11.02 (m, 1 H), 9.09 (m, 1 H), 8.74 (m, 1 H), 7.88 (m, 1 H), 7.67 (s, 1 H), 7.33

(m, 1 H), 7.27 (m, 1 H), 7.23 (m, 1 H), 7.12 (m, 1 H), 6.89 (m, 1 H), 5.94 (s, 2 H), 1.74 (m, 4 H).

Reference compound I-552

The title compound was prepared following general procedure C in the library format, except that thiazole-5-carboxylic acid was the acid reagent, and the crude material was purified by reverse phase HPLC providing the desired compound, the compound reference I-552 (2.3 mg, 9% yield) as a white solid.

1H-NMR (500 MHz, DMSO-d6) 5 ppm 11.71 (m, 1 H), 9.35 (m, 1 H), 9.12 (m, 1 H), 8.96 (m, 1 H), 8.79 (m, 1H), 8.08

(m, 1 H), 7.70 (m, 1 H), 7.34 (m, 1 H), 7.24 (m, 2 H), 7.12 (m, 1 H), 6.89 (m, 1 H), 5.95 (m, 2 H).

Reference compound I-553

The composition of the titles was prepared following the general management procedure in the library form, except for acid.

6 - o x o - 1 ,6 - d i h i d r o p i r i d i n - 2 - c a r b o x y l i c o was the acid reactive, and the raw material was purified by reverse phase H P L providing the desired compound, the reference compound I-553 (1 .9 mg, yield 7%) as a white solid.

Reference compound I-554

The title compound was prepared following general procedure C in the library format, except that 3-methoxyisoxazole-1-carboxylic acid was the acidic reagent, and the crude material was purified by reverse phase HPLC to provide the desired compound, the Reference compound I-554 (4.6 mg, 17% yield) as a white solid.

Reference compound I-555

The title compound was prepared following general procedure C in the library format, except that pyrimidine-4-carboxylic acid was the acidic reagent, and the crude material was purified by reverse phase HPLC providing the desired compound, the compound reference I-555 (1.6 mg, 6% yield) as a white solid.

Reference compound I-556

The title compound was prepared following general procedure C in the library format, except that oxazole-5-carboxylic acid was the acid reagent, and the crude material was purified by reverse phase HPLC providing the desired compound, the compound reference I-556 (4.4 mg, 17% yield) as a white solid.

Reference compound I-557

The title compound was prepared following general procedure C in the library format, except that oxazole-4-carboxylic acid was the acid reagent, and the crude material was purified by reverse phase HPLC providing the desired compound, the compound reference I-557 (4.4 mg, 17% yield) as a white solid.

Reference compound I-558

The title compound was prepared following general procedure C in the library format, except that cyclopropanecarboxylic acid was the acid reagent, and the crude material was purified by reverse phase HPLC providing the desired compound, the reference compound I-558 (5.1 mg, 21% yield) as a white solid.

1H-NMR (500 MHz, DMSO-da) 8 ppm 11.42 (m, 1 H), 9.11 (m, 1 H), 8.69 (m, 1 H), 8.01 (m, 1 H), 7.66 (m, 1 H), 7.34 (m, 1 H), 7.24 (m, 2 H), 7.12 (m, 1 H), 6.89 (m, 1 H), 5.93 (m, 2 H), 2.12 (m, 1 H), 0.87 (d, 4 H).

Reference compound I-559

The title compound was prepared following general procedure C in the library format, except that (S)-2-methoxy-2-phenylacetic acid was the acid reagent, and the crude material was purified by reverse phase HPLC providing the desired compound, reference compound I-559 (6.8 mg, 24% yield) as a white solid.

1H-NMR (500 MHz, DMSO-d6) 811.03 (m, 1 H), 9.09 (m, 1 H), 8.72 (m, 1 H), 7.96 (m, 1 H) , 7.67 (m, 1 H), 7.51 (m, 2 H), 7.36 (m, 4 H), 7.23 (m, 2 H), 7.12 (m, 1 H) , 6.87 (m, 1 H), 5.93 (m, 2 H), 5.12 (m, 1 H), 3.34 (s, 3 H).

Reference compound I-560

The title compound was prepared following general procedure C in the library format, except that furan-2-carboxylic acid was the acid reagent, and the crude material was purified by reverse phase HPLC providing the desired compound, the compound reference I-560 (5.2 mg, 20% yield) as a white solid.

1H-NMR (500 MHz, DMSO-d6) 8 ppm 11.10 (m, 1 H), 9.09 (m, 1 H), 8.75 (m, 1 H), 8.10 (m, 1 H), 8.00 (m, 1 H), 7.78 (m, 1 H), 7.69 (m, 1 H), 7.34 (m, 1 H), 7.24 (m, 2 H), 7.12 (m, 1 H), 6.89 (m, 1 H), 6.73 (m, 1 H), 5.94 (s, 2 H).

Reference compound I-561

The title compound was prepared following general procedure C in the library format, except that thiophen-2-carboxylic acid was the acid reagent, and the crude material was purified by reverse phase HPLC providing the desired compound, the compound reference I-561 (3.9 mg, 15% yield) as a white solid.

1H-NMR (500 MHz, DMSO-d6) 8 ppm 11.39 (m, 1 H), 9.10 (m, 1 H), 8.78 (m, 1 H), 8.36 (m, 1 H), 8.10 (m, 1 H), 7.95 (m, 1 H), 7.70 (s, 1 H), 7.34 (m, 1 H), 7.25 (m, 3 H), 7.12 (m, 1 H), 6.88 (m, 1 H), 5.95 (m, 2 H).

REFERENCE COMPOSITION I - 562

The composition of the titles was prepared following the general management procedure in the library form, except for acid.

2 - e t o x i a c e was the reactive acid, and the raw material was purified by means of HPL If this is the case, the reference compound I-562 (5.7 mg, yield 23%) as a solid n c o .

1H-NMR (500 MHz, DMSO-da) 8 ppm 10.70 (m, 1 H), 9.10 (m, 1 H), 8.73 (m, 1 H), 8.00 (m, 1 H), 7.66 (m, 1 H), 7, (m, 1 H), 7.23 (m, 2 H), 7.11 (m, 6.89 (m, 1 H), 5, 93 (s, 2 H), 4.17 (s, 2 H), 3.55 (m, 2 H), 1.17 (m, 3 H).

Reference compound I-563

The title compound was prepared following general procedure C in the library format, except that the acid

2-(methylsulfonyl)acetic acid was the acid reagent, and the crude material was purified by reverse phase HPLC to provide the desired compound, reference compound I-563 (3 mg, 11% yield) as a white solid.

1H-NMR (500 MHz, DMSO-da) 8 ppm 11.53 (m, 1 H), 9.08 (m, 1 H), 8.78 (m, 1 H), 7.99 (m, 1 H), 7.66 (m, 1 H), 7, (m, 1 H), 7.23 (m, 2 H), 7.12 (m, 6.91 (m, 1 H), 5, 93 (m, 2 H), 4.46 (m, 2 H), 3.17 (s, 3 H).

Reference compound I-564

The title compound was prepared following general procedure C in the library format, except that the acid

3-Cyclopropyl-1H-pyrazole-5-carboxylic acid was the acidic reagent, and the crude material was purified by reverse phase HPLc providing the desired compound, reference compound I-564 (1.2 mg, 4% yield) as a white solid.

1H-NMR (500 MHz, DMSO-d6) 8 ppm 9.84 (br s, 1 H), 9.10 (m, 1 H), 8.77 (m, 1 H), 8.07 (m, 1H), 7.72 (m, 1H), 7.34

(m, 1 H), 7.28 (m, 1 H), 7.23 (m, 1 H), 7.12 (m, 1 H), 6.89 (m, 1 H), 6.61 (m, 1 H), 5.95 (m, 2 H), 1.96 (m, 1 H), 0.98 (m,

2H), 0.76 (m, 2H).

Reference compound I-565

The title compound was prepared following general procedure C in the library format, except that the acid

2-acetoxy-2-phenylacetic acid was the acid reagent, and the crude material was purified by reverse phase HPLC to provide the desired compound, reference compound I-565 (4.1 mg, 14% yield) as a white solid.

1H-NMR (500 MHz, DMSO-d6) 8 ppm 11.60 (m, 1 H), 9.08 (m, 1 H), 8.72 (m, 1 H), 7.92 (m, 1 H), 7.65 (m, 1H), 7.59

(m, 2 H), 7.41 (m, 3 H), 7.33 (m, 1 H), 7.23 (m, 2 H), 7.11 (m, 1 H), 6.88 (m, 1 H), 6.17 (s, 1 H), 5.92 (s, 2 H), 2.15 (s,

3H).

Reference compound I-569

The title compound was prepared following general procedure C in the library format, except that the acid

1-methylcyclopropane-1-carboxylic acid was the acid reagent, and the crude material was purified by reverse phase HPLC to provide the desired compound, reference compound I-569 as a white solid.

1H-NMR (500 MHz, CD3OD) 8 ppm 8.81 (m, 1 H), 8.69 (m, 1 H), 8.18 (m, 1 H), 7.59 (m, 1 H) , 7.31 (m, 1 H), 7.10 (m,

2 H), 6.93 (m, 2 H), 6.01 (m, 2 H), 1.52 (s, 3 H), 1.32 (m, 2 H), 0.84 (m, 2H).

Reference compound I-570

The title compound was prepared following general procedure C in the library format, except that tetrahydrofuran-2-carboxylic acid was the acid reagent, and the crude material was purified by reverse phase HPLC providing the desired compound, the compound reference I-570 as a white solid.12

1H-NMR (500 MHz, CD3OD) 8 ppm 8.79 (m, 1 H), 8.72 (m, 1 H), 8.17 (m, 1 H), 7.55 (m, 1 H) , 7.29 (m, 1 H), 7.09 (m, 2 H), 6.92 (m, 2 H), 5.99 (m, 2 H), 4.53 (m, 1 H) , 4.13 (m, 1 H), 3.98 (m, 1 H), 2.39 (m, 1 H), 2.14 (m, 1 H), 2.01 (m, 2

H).

Reference compound I-571

The title compound was prepared following general procedure C in the library format, except that the acid

2-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)acetic acid was the acid reagent, and the crude material was purified by reverse phase HPLC to provide the desired compound, compound reference I-571 as a white solid.

1 H - R M N (500 M H z, C D 3 O D) 8 p p m 8, 79 (d, 1 H), 8, 69 (d, 1 H), 8, 09 (d, 1 H), 7, 54 (s , 1 H ) , 7 , 43 ( d , 1 H ) , 7 , 3 2 - 7 , 25 ( m ,

1 H), 7, 14 - 7, 0 1 (m, 2 H), 6, 96 - 6, 89 (m, 1 H), 6, 88 (d, 1 H), 5, 98 (s, 2 H), 4, 70 (s, 2 H), 1, 91 (d, 3 H).

REFERENCE COMPOSITION I - 572

The composition of the titles was prepared following the general management procedure in the library format, except for the acid 3, 3, 3 - tri fl uor o - 2 - m e t o x i - 2 - f e n i l p r o p ano i c o was the acidic reagent, and the raw material was purified by means of H P L C of phase and in v e r s a p p r o p o r t i o n a n d t h e c o m p u e s t o d e a d e , t h e c o m p u e s t o r e f e r e n c i a - 572 as a b l a n c or .

1H-NMR (500 MHz, CD3OD) 88.79 (m, 2 H), 8.19 (m, 1 H), 7.64 (m, 2 H), 7.57 (m, 1 H), 7 .48 (m, 3 H), 7.30 (m, 1 H), 7.08 (m, 2 H), 6.92 (m, 2 H), 5.99 (m, 2 H), 3 .63 (d, 3H).

Reference compound I-574

The title compound was prepared following general procedure C in the library format, except that tetrahydro-2H-pyran-4-carboxylic acid was the acid reagent, and the crude material was purified by reverse phase HPLC to provide the desired compound. , the reference compound I-574 as a white solid.

1H-NMR (500 MHz, CD3OD) 8 ppm 8.81 (d, 1 H), 8.66 (s, 1 H), 8.20 (d, 1 H), 7.54 (s, 1 H) , 7.30 (m, 1 H), 7.09 (m, 2 H), 6.93 (m, 1 H), 6.88 (d, 1 H), 5.98 (s, 2 H) , 4.02 (m, 2 H), 3.52 (m, 2 H), 2.80 (m, 1 H), 1.85 (d, 4 H).

Reference compound I-577

The acetyl-protected intermediate was prepared following general procedure C in the library format, except that 2-acetoxybenzoic acid was the acid reagent. The crude material was purified by reverse phase HPLC providing the desired intermediate product. The intermediate product was then dissolved in a mixture of methanol:water (8:1) and treated with lithium hydroxide (4.5 equiv.) and a small amount of THF (300 μl). After completing the reaction, the volatile substances were removed in vacuo and the residue was treated with 1N HCl solution until reaching pH ~4. The mixture was extracted with ethyl acetate and the organic layers were washed with water and brine. The contents were dried with sodium sulfate, filtered and concentrated to provide the desired compound, reference compound I-577 (10 mg, 33% yield, 2 steps) as a white solid.

1H-NMR (500 MHz, DMSO-d6) 8 ppm 11.81 (m, 1 H), 11.12 (m, 1 H), 9.10 (d, 1 H), 8.79 (m, 1 H), 8.20 (m, 1 H), 8.00 (m, 1 H), 7.66 (s, 1 H), 7.51 (m, 1 H), 7.34 (m, 1 H), 7.28 (d, 1 H), 7.23 (m, 1 H), 7.08 (m, 3 H), 6.90 (m, 1 H), 5.96 (s, 2 H).

Reference compound I-579

A solution of 2-cyanoacetic acid (4 equiv.) in DMF was cooled to 0 °C, and treated with oxalyl chloride (4.1 equiv.) as a solution in DMF. Gas release was observed and the contents were stirred at 0 °C for 30 min. Intermediate 2 (1 equiv.) was added to the reaction, and the contents were stirred for 18 h, while allowing to warm to 23 °C. The solvents were removed in vacuo and the crude material was purified by reverse phase HPLC to provide the desired compound, reference compound I-579 (2.3 mg, 10% yield) as a white solid.

1H-NMR (500 MHz, DMSO-d6) 8 ppm 11.47 (m, 1 H), 9.10 (d, 1 H), 8.78 (m, 1 H), 7.94 (m, 1 H), 7.65 (s, 1 H), 7.34 (d, 1 H), 7.24 (m, 2 H), 7.12 (m, 1 H), 6.91 (t, 1 H), 5.92 (s, 2 H), 4.05 (s, 2 H).

Reference compound I-594

The title compound was prepared following general procedure C in the library format, except that 2-methyl-2,3-dihydrobenzo[b]thiophen-2-carboxylic acid 1,1-dioxide was the acid reagent and the Crude material was purified by reverse phase HPLC providing the desired compound, reference compound I-594 (7.4 mg, 23% yield) as a yellow solid.

1H-NMR (500 MHz, DMSO-d6) 8 ppm 10.96 (m, 1 H), 9.09 (m, 1 H), 8.77 (m, 1 H), 7.93 (m, 1 H), 7.79 (m, 1 H), 7.68 (s, 2 H), 7.57 (m, 2 H), 7.34 (m, 1 H), 7.24 (m, 2 H), 7.12 (m, 1 H), 6.89 (m, 1 H), 5.94 (s, 2 H), 4.11 (m, 1 H), 3.30 (m, 1 H), 1.89 (s, 3 H).

Reference compound I-596

The title compound was prepared following general procedure C in the library format, except that 2-(1,3-dioxoisoindolin-2-yl)acetic acid was the acid reagent, and the crude material was purified by phase HPLC. reverse providing the desired compound, reference compound I-596 (17.4 mg, 56% yield) as a white solid.

1H-NMR (500 MHz, DMSO-d6) 8 ppm 11.60 (m, 1 H), 9.09 (m, 1 H), 8.74 (m, 1 H), 7.91 (m, 5 H), 7.65 (m, 1 H), 7.35 (m, 1 H), 7.24 (m, 2 H), 7.12 (m, 1 H), 6.92 (m, 1 H), 5.93 (m, 2 H), 4.58 (s, 2 H).

Reference compound I-597

The composition of the titles was prepared following the general management procedure in the library format, except for the l) glycine was the acid reagent, and the raw material was purified by reverse-phase HPLC by proportioning the component. s e a d , the c o m p u e s t o d e f e r e n c e I-597 (4.4 mg, yield 15%) as a white solid.

1 H - R M N ( 500 M H z , C D 3O D ) 5 p p m 8 , 83 ( m , 1 H ) , 8 , 68 ( m , 1 H ) , 8 , 24 ( m , 1 H ) , 7 , 58 ( s , 1 H ), 7 , 34 ( m , 5 H ) , 7 , 26 ( m , 1 H ) , 7 , 10 ( m , 2 H ) , 6 .97 ( m , 1 H ) , 6 .90 ( m , 1 H ), 6 , 01 ( s , 2 H ) , 4 , 17 ( s , 2 H ) , 3 , 66 ( s , 2 H ) .

Reference compound I-598

The title compound was prepared following general procedure C in the library format, except that (benzyloxy)carbonyl)glycine was the acid reagent, and the crude material was purified by reverse phase HPLC providing the desired compound, the compound reference I-598 (4 mg, 13% yield) as a white solid.

1H-NMR (500 MHz, DMSO-d6) 5 ppm 11.15 (m, 1 H), 9.10 (d, 1 H), 8.73 (m, 1 H), 7.98 (m, 1 H), 7.65 (s, 1 H), 7.57 (m, 1 H), 7.37 (m, 6 H), 7.23 (m, 2 H), 7.12 (m, 1 H), 6.90 (m, 1 H), 5.92 (m, 2 H), 5.05 (s, 2 H), 3.93 (m, 2 H).

Reference compound I-599

The title compound was prepared following general procedure C in the library format, except that 2-(1-dioxoisoindolin-2-yl)acetic acid was the acid reagent, and the crude material was purified by reverse phase HPLC to provide the desired compound, reference compound I-599 (11.7 mg, 39% yield) as a white solid.

1H-NMR (500 MHz, DMSO-d6) 5 ppm 11.42 (m, 1 H), 9.10 (m, 1 H), 8.74 (m, 1 H), 7.95 (m, 1 H), 7.72 (m, 1 H), 7.66 (m, 1 H), 7.63 (m, 2 H), 7.52 (m, 1 H), 7.34 (m, 1 H), 7.24 (m, 2 H), 7.13 (m, 1 H), 6.92 (m, 1 H), 5.93 (m, 2 H), 4.55 (m, 4 H).

Reference compound I-610

The title compound was prepared following general procedure C in the library format, except that 2-(2-oxooxazolidin-3-yl)acetic acid was the acid reagent, and the crude material was purified by reverse phase HPLC to provide the desired compound, reference compound I-610 (11.4 mg, 42% yield) as a white solid.

1H-NMR (500 MHz, DMSO-d6) 5 ppm 11.33 (m, 1 H), 9.10 (m, 1 H), 8.75 (m, 1 H), 7.96 (m, 1 H), 7.64 (m, 1 H), 7.35 (m, 1 H), 7.24 (m, 2 H), 7.12 (m, 1 H), 6.91 (m, 1 H), 5.93 (m, 2 H), 4.33 (m, 2 H), 4.15 (s, 2 H), 3.64 (m, 2 H).

Reference compound I-601

The title compound was prepared following general procedure C in the library format, except that 2-(4-oxoquinazolin-3(4H)-yl)acetic acid was the acidic reagent, and the crude material was purified by HPLC. reverse phase providing the desired compound, reference compound I-601 (3.3 mg, 11% yield) as a white solid.

1H-NMR (500 MHz, DMSO-d6) 5 ppm 11.69 (m, 1 H), 9.10 (m, 1 H), 8.75 (m, 1 H), 8.37 (s, 1 H), 8.15 (m, 1 H), 7.90 (m, 2 H), 7.73 (m, 1 H), 7.67 (s, 1 H), 7.56 (m, 1 H), 7.34 (m, 1 H), 7.24 (m, 2 H), 7.13 (m, 1 H), 6.93 (m, 1 H), 5.94 (s, 2 H), 4.99 (s, 2H).

Reference compound I-602

The title compound was prepared following general procedure C in the library format, except that (2-(1,3-dioxoisoindolin-2-yl)acetyl)glycine was the acid reagent, and the crude material was purified by HPLC reverse phase providing the desired compound, reference compound I-602 (1.2 mg, 4% yield) as a white solid.

1H-NMR (500 MHz, DMSO-d6) 5 ppm 11.20 (m, 1 H), 9.09 (m, 1 H), 8.74 (m, 1 H), 8.62 (m, 1 H), 7.98 (m, 1 H), 7.91 (s, 2 H), 7.88 (s, 2 H), 7.64 (s, 1 H), 7.33 (m, 1 H), 7.24 (d, 1 H), 7.20 (m, 1 H), 7.11 (m, 1 H), 6.89 (m, 1 H), 5.92 (m, 2 H), 4.29 (s, 2 H), 4.05 (m, 2 H).

Reference compound I-603

The title compound was prepared following general procedure C in the library format, except that (methoxycarbonyl)glycine was the acid reagent, and the crude material was purified by reverse phase HPLC providing the desired compound, reference compound I -603 (2.2 mg, 8% yield) as a white solid.

H), 7, 42

REFERENCE COMPOSITION I - 592

1H-NMR (500 MHz, DMSO-da) 8 ppm 11.44 (m, 1 H), 9.10 (m, 1 H), 8.75 (m, 1 H), 7.92 (m, 1 H), 7.91 (m, 1 H), 7.89 (m, 1 H), 7.88 (m, 1 H), 7.76 (m, 1 H), 7.65 (m, 2 H), 7.34 (m, 1 H), 7.26 (m, 1 H), 7.22 (m, 1 H), 7.12 (m, 1 H), 6.91 (m, 1 H), 5.92 (m, 2 H), 4.67 (m, 2 H).

Reference compound I-594

The title compound was prepared following general procedure C except that 2-((4-chlorophenyl)sulfonyl)acetic acid was the acid reagent, and the crude material was purified by reverse phase HPLC providing the desired compound, the compound reference I-594 (5.8 mg, 15% yield) as a white solid.

1H-NMR (500 MHz, DMSO-d6) 8 ppm 11.48 (s, 1 H), 9.10 (d, 1 H), 8.76 (d, 1 H), 7.92 (m, 2 H), 7.87 (m, 1 H), 7.76 (m, 2 H), 7.65 (s, 1 H), 7.34 (m, 1 H), 7.24 (m, 2 H), 7.12 (m, 1 H), 6.91 (m, 1 H), 5.93 (s, 2 H), 4.73 (m, 2 H).

Reference compound I-498

To a mixture of 5-fluoro-2-(1-(2-fluorobenzyl)-5-(isoxazol-3-yl)-1H-pyrazol-3-yl)--pyrimidin-4-amine (intermediate product described in WO2012 /3405 A1; 1 equiv.), triethylamine (6 equiv.), and N,N-dimethylpyridin-4-amine (0.01 equiv.) in dichloromethane 2-chloro-2-oxoethyl acetate (3 equiv. ) at 23 °C. The contents were heated to 60 °C and stirred for 18 h. The solvent was removed in vacuo and purification of the crude material by reverse phase HPLC provided the desired compound, reference compound I 498 (1.0 mg, 2% yield) as a solid.

1H-NMR (500 MHz, DMSO-d6) 8 ppm 11.21 (m, 1 H), 9.09 (m, 1 H), 8.84 (m, 1 H), 7.61 (m, 1 H), 7.34 (m, 1 H), 7.27 (m, 1 H), 7.22 (m, 1 H), 7.12 (m, 1 H), 6.94 (m, 1 H), 5.92 (m, 2 H), 4.91 (s, 2 H), 2.13 (s, 3 H).

Reference compound I-578

The title compound was prepared following general procedure B, except that tetrahydrofuran-3-amine was the amine reagent, 6 equivalents of triethylamine were used, and the contents were heated to 100 °C as a solution in dioxane/water ( 4:1) for 24 hours. The mixture was cooled to 23°C and the solvent was removed in vacuo. The solid was purified by reverse phase HPLC to provide the desired compound, reference compound I-578 (12 mg, 53% yield) as a solid.

1H-NMR (500 MHz, DMSO-d6) 8 ppm 9.11 (m, 1 H), 8.58 (m, 1 H), 8.31 (m, 1 H), 7.61 (m, 1 H), 7.34 (m, 1 H), 7.24 (m, 2 H), 7.11 (m, 1 H), 6.89 (m, 1 H), 5.93 (s, 2 H), 4.76 (m, 1 H), 4.04 (m, 1 H), 3.89 (m, 1 H), 3.77 (m, 1 H), 3.64 (m, 1 H), 2.27 (m, 1 H), 2.04 (s, 1 H).

Reference compound I-613

A solution of intermediate 1 (1 equiv.) in DMSO was treated with potassium benzenesulfonamide (2 equiv.).

The resulting reaction mixture was stirred at 100 °C for 8 h. The content was filtered, and the filtrate was directly purified by reverse phase HPLC to provide the desired compound, reference compound I-613 (7 mg, 26% yield) as a solid.

1H-NMR (500 MHz, CD3OD) 8 ppm 8.82 (m, 1 H), 8.37 (m, 1 H), 8.26 (m, 2 H), 7.58 (m, 1 H) , 7.47 (m, 2 H), 7.31 (m, 2 H), 7.11 (m, 2H), 6.94 (m, 2 H), 5.98 (m, 2 H).

Reference compound I-614

A solution of intermediate 1 (1 equiv.) in DMSO was treated with potassium 3,4-dimethoxybenzenesulfonamide (2 equiv.). The resulting reaction mixture was stirred at 100 °C for 8 h. The content was filtered, and the filtrate was directly purified by reverse phase HPLC to provide the desired compound, reference compound I-614 (1.3 mg, 5% yield) as a solid.12

1H-NMR (500 MHz, DMSO-d6) 8 ppm 9.12 (m, 1 H), 8.56 (m, 1 H), 7.90 (m, 1 H), 7.60 (m, 1 H), 7.38 (m, 2 H), 7.27 (d, 2 H), 7.12 (m, 1 H), 6.92 (m, 2 H), 5.94 (s, 2 H), 3.71 (d, 6 H).

Reference compound I-607

A solution of intermediate 1 (1 equiv.) in DMSO was treated with potassium (4-fluorophenyl)methanesulfonamide (2 equiv.). The resulting reaction mixture was stirred at 60 °C for 0.5 h, after which the reaction was diluted with water and 1N hydrochloric acid solution and extracted with dichloromethane. The combined organic layers were dried over sodium sulfate, filtered, and concentrated in vacuo. The crude material was purified by silica gel chromatography using a gradient of 0-5% methanol in dichloromethane providing the desired compound, reference compound I-607 (2.8 mg, 6% yield) as a solid.

1 H - R M N ( 500 MHz , C D 3 O D ) 5 p p m 8 , 79 ( m , 1 H ) , 8 , 45 ( m , 1 H ) , 7 , 52 ( m , 1 H ) , 7 , 40 ( m , 2 H), 7, 26 (m, 1 H), 6, 94 (d, 6 H), 5, 98 (s, 2 H), 5, 04 (m, 2 H).

Reference compound I-624

To a solution of 4-fluorobenzenesulfonamide (4 equiv.) in DMF was added potassium bis(trimethylsilyl)amide (4 equiv.) at 23 °C. After 15 minutes of stirring, intermediate 1 (1 equiv.) was added and the reaction was stirred for 3 days at 75 °C. Raw, the product was purified by reverse phase HPLC to provide the desired compound, reference compound I-624 (1.9 mg, 7% yield) as a solid.

1H-NMR (500 MHz, DMSO-da) 5 ppm 9.13 (m, 1 H), 8.51 (m, 1 H), 8.30 (m, 2 H), 7.40 (m, 2 H), 7.27 (m, 2 H), 7.15 (m, 3 H), 6.95 (m, 1 H), 5.99 (s, 2 H).

Reference compound I-625

A solution of intermediate 1 (1 equiv.) in DMSO was treated with potassium pyridine-3-sulfonamide (1 equiv.) and potassium carbonate (0.5 equiv.). The resulting reaction mixture was heated at 150 °C for 10 min. in a microwave. The content was filtered, and the filtrate was directly purified by reverse phase HPLC to provide the desired compound, reference compound I-625 (4.4 mg, 33% yield) as a solid.

1H-NMR (500 MHz, DMSO-d6) 5 ppm 9.24 (m, 1 H), 9.14 (m, 1 H), 8.69 (m, 1 H), 8.59 (m, 1 H), 8.40 (m, 1 H), 7.42 (m, 1 H), 7.34 (m, 2 H), 7.24 (m, 2 H), 7.13 (m, 1 H), 6.96 (m, 1 H), 5.95 (m, 2 H).

Reference compound I-583

A solution of intermediate 2 (1 equiv.) in DMF was treated with isocyanatobenzene (2 equiv.) and triethylamine (2 equiv.). The resulting reaction mixture was heated at 100 °C for 18 h. The content was filtered, and the filtrate was directly purified by reverse phase HPLC to provide the desired compound, reference compound I-583 (1.0 mg, 4% yield) as a solid. 1H-NMR (500 MHz, DMSO-d6) 5 ppm 10.20 (m, 1 H), 9.10 (m, 1 H), 8.62 (m, 1 H), 7.71 (m, 1 H), 7.63 (m, 2 H), 7.27 (m, 7 H), 7.09 (m, 1 H), 7.01 (m, 1 H), 6.88 (m, 1 H), 5.99 (m, 2H).

Reference compound I-491

To a solution of intermediate 2 (1 equiv.) in dichloromethane was added (4-fluorophenyl)methanesulfonyl chloride (1 equiv.), followed by DBU (1 equiv.). The reaction was stirred at 90 °C for 18 h. The reaction mixture was diluted with water, extracted with dichloromethane (3 times), washed with 1N hydrochloric acid solution (2 times), dried (sodium sulfate), filtered and concentrated in vacuo. Purification of the crude material by reverse phase HPLC provided the desired compound, reference compound I-491 (17.8 mg, 17% yield) as a solid.

1H NMR (500 MHz, CDCh) 5 ppm 8.52 (d, 1 H), 8.28 (br. s., 1 H), 7.38 (br. s., 1 H), 7.30 ( dd, 2 H), 7.25 (br. s., 1 H), 7.14-6.97 (m, 4 H), 6.92-6.73 (m, 3 H), 6.63 (d, 1 H), 5.91 (s, 2 H), 4.54 (br. s., 2 H).

Reference compound I-495

The title compound was prepared following general procedure B, except that ethanol-1,1,2,2-d4-amine was the amine reagent, and the contents were heated at 90 °C for 20 h. The content was cooled to 23 °C, and was distributed between a 1:1 mixture of dichloromethane and 1N HCl solution. The layers were separated and the aqueous layer was extracted with dichloromethane (2 times), and the organic parts were combined and washed with brine. The mixture was dried with magnesium sulfate, filtered and concentrated in vacuo. The crude material was purified by silica gel chromatography using a gradient of 0-10% methanol in dichloromethane providing the desired compound, reference compound I-495 (120 mg, 74% yield) as a solid.

1H NMR (500 MHz, DMSO-d6) 5 ppm 9.09 (d, 1 H), 8.17 (d, 1 H), 7.61 (s, 1 H), 7.51 (s, 1 H) ), 7.33 (d, 1 H), 7.14 - 7.28 (m, 2 H), 7.10 (t, 1 H), 6.82 (s, 1 H), 5.90 ( s, 2 H), 4.74 (br. s., 1 H).

Reference compound I-505

5-fluoro-2-(1-(2-fluorobenzyl)-5-(isoxazol-3-yl)-1H-pyrazol-3-yl)pyrimidin-4-amine (intermediate product described in WO2012/3405 A1) was added; 1 equiv.) to a suspension of NaH (1.2 equiv.) in anhydrous THF at 23 °C. After stirring for 30 minutes at 23 °C, a solution of propane-2-sulfonyl chloride (1 equiv.) in THF was added to the reaction mixture. The contents were heated to 70 °C and stirred for an additional 18 h. The reaction mixture was diluted with water, extracted with dichloromethane (3 times), dried (sodium sulfate), filtered and concentrated in vacuo. Purification of the crude material by reverse phase HPLC provided the desired compound, reference compound I-505 (2.9 mg, 6% yield) as a solid.

1 H R M N ( 500 M H z , C D C l a ) 5 p p m 11 , 59 ( b r . s . , 1 H ) , 9 , 13 ( d , 1 H ) , 8 , 66 ( br . s . , 1 H ) , 7 , 45 (s, 1H), 7, 3 8 - 7, 3 1 (m, 1H), 7, 26 (d, 1H), 7, 24 - 7, 17 (m, 1H), 7, 13 (t, 1 H), 7, 0 8 - 7, 0 2 (m, 1 H), 5, 89 (s, 2 H), 4, 24 (b r. s., 1 H), 1, 35 (d, 6H).

REFERENCE COMPOSITION I - 510

To a solution of intermediate 2 (1 equiv.) in dichloromethane was added DBU (1 equiv.) followed by methyl 2-(chlorosulfonyl)acetate (1 equiv.). The reaction was stirred at 90 °C for 18 h. The reaction mixture was diluted with water, extracted with dichloromethane (3 times), washed with 1N hydrochloric acid solution (2 times), dried (sodium sulfate), filtered and concentrated in vacuo. Purification of the crude material by reverse phase HPLC provided the desired compound, reference compound I-510 (8.3 mg, 12% yield) as a solid.

1H NMR (500 MHz, CDCh) 8 ppm 8.52 (d, 1 H), 8.37 (d, 1 H), 7.40 (s, 1 H), 7.26-7.22 (m, 1 H), 7.08-6.99 (m, 4 H), 6.63 (d, 1 H), 5.93 (s, 2 H), 4.31 (s, 2 H), 3, 71 (s, 3 H).

Reference compound I-521

The title compound was prepared following general procedure B, except that 3-amino-2,2-difluoropropan-1-ol (1.5 equiv., as the HCl salt) was the amine reagent, 1 equivalent of triethylamine, and the contents were heated at 60 °C for 20 h. The content was cooled to 23 °C, and was distributed between a 1:1 mixture of dichloromethane and 1N HCl solution. The layers were separated and the aqueous layer was extracted with dichloromethane (2 times), and the organic parts were combined and washed with brine. The mixture was dried over Na2SO4, filtered and concentrated in vacuo. The crude material was purified by silica gel chromatography using a gradient of 0-10% methanol in dichloromethane providing the desired compound, reference compound I-521 (36 mg, 60% yield) as a solid.

1H-NMR (500 MHz, CDCla) 8 ppm 8.48 (d, 1 H), 8.32 (d, 1 H), 7.42 (s, 1 H), 7.24-7.19 (m , 1 H), 7.04-6.97 (m, 3 H), 6.60 (d, 1 H), 6.05 (br. s., 1 H), 5.93 (s, 2 H) ), 4.12 (td, 2 H), 3.74 (t, 2 H).

Reference compound I-539

To a solution of the reference compound I-510 (1 equiv.) in THF was added sodium borohydride (3 equiv.) at 23 °C. The reaction mixture was heated to 75 °C, methanol (4 equiv.) was added dropwise with a syringe, and the contents were stirred for 1 h. After cooling to 23 °C, the reaction was concentrated in vacuo and the resulting crude material was purified by reverse phase HPLC to provide the desired compound, reference compound I-539 (1.5 mg, 27% yield) as a solid.

1H NMR (500 MHz, CD3OD) 8 ppm 8.69 (d, 1 H), 8.35 (d, 1 H), 7.47 (s, 1 H), 7.19 (d, 1 H), 6.96 (s, 3 H), 6.89 (d, 1 H), 6.84-6.82 (m, 1 H), 5.89 (s, 2 H), 3.92 (t, 2H), 3.66 (t, 2H).

Reference compound I-610

A solution of intermediate 1 (1 equiv.) in DMSO was treated with potassium (cyclopropylsulfonyl)amide (2 equiv.). The resulting reaction mixture was stirred at 23 °C for 16 h. The content was filtered, and the filtrate was directly purified by reverse phase HPLC to provide the desired compound, reference compound I-610 (34 mg, 55% yield) as a solid. 1H-NMR (500 MHz, CD3OD) 8 ppm 8.66 (d, 1 H), 8.35 (d, 1 H), 7.37 (s, 1 H), 7.23-7.11 (m , 1 H), 7.02-6.89 (m, 2 H), 6.86-6.74 (m, 2 H), 5.91-5.77 (m, 2 H), 3.36 -3.26 (m, 1 H), 1.35 1.17 (m, 2 H), 1.08-0.89 (m, 2 H).

Reference compound I-611

A solution of intermediate 1 (1 equiv.) in DMSO was treated with potassium (propylsulfonyl)amide (2 equiv.). The resulting reaction mixture was stirred at 23 °C for 16 h. The content was filtered, and the filtrate was directly purified by reverse phase HPLC to provide the desired compound, reference compound I-611 (50 mg, 81% yield) as a solid. 1H-NMR (500 MHz, CD3OD) 8 ppm 8.67 (d, 1 H), 8.36 (d, 1 H), 7.33 (s, 1 H), 7.23 7.13 (m, 1 H), 7.03-6.89 (m, 2 H), 6.87-6.81 (m, 1 H), 6.79 (d, 1 H), 5.91-5.68 ( m, 2 H), 3.63 (t, 2 H), 1.93-1.74 (m, 2 H), 0.97 (t, 3 H).

Reference compound I-629

A solution of intermediate 1 (1 equiv.) in DMSO was treated with potassium methyl(methylsulfonyl)amide (1 equiv.). The resulting reaction mixture was stirred at 23 °C for 16 h. The content was filtered, and the filtrate was directly purified by reverse phase HPLC to provide the desired compound, reference compound I-629 (8 mg, 33% yield) as a solid.

1H-NMR (500 MHz, DMSO-d6) 8 ppm 9.11 (d, 1 H), 8.93 (d, 1 H), 7.62 (s, 1 H), 7.34 (d, 1 H), 7.28 (d, 1 H), 7.24-7.19 (m, 1 H), 7.12 (td, 1 H), 6.94 (td, 1 H), 5.92 (s, 2 H), 3.49 (s, 3 H), 3.37 (d, 3 H).

Reference compound I-475

The title compound was prepared following general procedure C, except that (S)-2-acetoxy-3,3,3-trifluoropropanoic acid was the acid reagent (3 equiv.), 7 equivalents of triethylamine were used, and they used 4 equivalents of T3P. The solution was heated to 50 °C for 10 minutes, at which time the solution was diluted with ethyl acetate and water. The layers were separated and the organic layer was washed with water and saturated aqueous sodium chloride. The organic substances were dried over magnesium sulfate, filtered, and the solvent was removed in vacuo. P u r i f i c a t i o n m e d i a n t e ch r o m a t o g r a p h y e n si l i c e (grade 0 - 5% m e t a n o l e n d i c h l o r o m e t h a n e ) p r o p o r t i o n t h e c o As a result, the reference compound I-475 (98 mg, quantitative yield) as a solid o b la n c o .

1H-NMR (500 MHz, CDCh) 8 ppm 8.81 (d, 1 H), 8.78 (s, 1 H), 8.50 (d, 1 H), 8.15 (d, 1 H) , 7.51 (s, 1 H), 7.25-7.22 (m, 1 H), 7.09-7.05 (m, 1 H), 7.00 (t, 1 H), 6 .85-6.82 (m, 1 H), 6.63 (d, 1 H), 6.06 (s, 2 H), 5.77 (q, 1 H), 2.39 (s, 3 H).

Reference compound I-485

The title compound was prepared following general procedure B, except that 4-aminopyrrolidin-2-one was the amine reagent. After stirring at 90 °C for 16 h, additional water was added to solubilize the reagents. After 5 h, the reaction mixture was partitioned between dichloromethane and saturated aqueous sodium bicarbonate. The layers were separated and the aqueous layer was extracted with dichloromethane. The organic substances were dried over magnesium sulfate, filtered, and the solvent was removed in vacuo. Purification by silica gel chromatography (0-15% methanol in dichloromethane gradient) provided the desired compound, reference compound I-485 (11 mg, 19% yield) as a white solid.

1H-NMR (500 MHz, CD3OD) 8 ppm 8.76 (s, 1 H), 8.11 (d, 1 H), 7.46 (s, 1 H), 7.27 (q, 1 H) , 7.11-7.07 (m, 1 H), 7.03 (t, 1 H), 6.93 (s, 1 H), 6.83-6.80 (m, 1 H), 5 .96 (s, 2 H), 5.11-5.06 (m, 1 H), 3.90 (dd, 1 H), 3.40 (dd, 1 H), 2.84 (dd, 1 H), 2.51 (dd, 1 H).

Reference compound I-500

A solution of 2-(1-(2-fluorobenzyl)-5-(isoxazol-3-yl)-1H-pyrazol-3-yl)pyrimidin-4-ol (intermediate product described in WO2012/003405 A1; 1 equiv. ) and phosphoryl trichloride (20 equiv.) was heated at 60 °C for 1 h, after which the phosphoryl trichloride was removed in vacuo. The resulting residue was dissolved in dioxane and water (2:1 ratio). 2-(aminomethyl)-1,1,1,3,3,3-hexafluoropropan-2-ol (3 equiv.) and triethylamine (10 equiv.) were added and the resulting solution was heated at 110 °C for 7 d. . The solution was partitioned between 1 N aqueous hydrochloric acid and dichloromethane. The aqueous layer was extracted with dichloromethane. The organic substances were dried over magnesium sulfate, filtered, and the solvent was removed in vacuo. Purification by silica gel chromatography (0-100% gradient (7:1 = acetonitrile:methanol) in dichloromethane) provided the desired compound, reference compound I-500 (16 mg, 21% yield) as a solid. yellow.

1H-NMR (500 MHz, CD3OD) 8 ppm 8.77 (s, 1 H), 8.20 (d, 1 H), 7.37 (s, 1 H), 7.29-7.24 (m , 1 H), 7.09-7.00 (m, 3 H), 6.85 (s, 1 H), 6.61 (d, 1 H), 5.95 (s, 2 H), 4 .08 (s, 2 H).

Reference compound I-518

The title compound was prepared following general procedure B, except that 3,3,3-trifluoropropane-1,2-diamine (9 equiv.) was the amine reagent, and 30 equivalents of triethylamine were used. After stirring for 16 h, the crude reaction mixture was partitioned between water and ethyl acetate. The aqueous layer was extracted with ethyl acetate. The organic substances were dried over magnesium sulfate, filtered, and the solvent was removed in vacuo. Purification by silica gel chromatography (0-5% methanol in dichloromethane gradient) provided the desired compound, reference compound I-518 (10 mg, 21% yield) as a white solid.

1H-NMR (500 MHz, CD3OD) 8 ppm 8.39 (d, 1 H), 8.11 (d, 1 H), 7.19 (s, 1 H), 7.14-7.10 (m , 1 H), 6.97-6.93 (m, 1 H), 6.89 (t, 1 H), 6.81-6.78 (m, 1 H), 6.52 (d, 1 H), 5.90 (s, 2 H), 5.57 (br s, 1 H), 4.17 (ddd, 1 H), 3.51-3.44 (m, 1 H), 3, 34 (ddd, 1 H).

Reference compound I-540

A solution of reference compound I-403 (1 equiv.) in dichloromethane was treated with diisopropylethylamine (2 equiv.), followed by HATU (1.5 equiv.). After stirring for 20 minutes, ammonia (3 equiv., 0.5 M solution in dioxane) was added. After 22 h, additional ammonia (3 equiv.) was added. After 4 hours, the solution was diluted with aqueous 1N hydrochloric acid solution and dichloromethane. The layers were separated and the aqueous layer was extracted with dichloromethane. The organic substances were dried over magnesium sulfate, filtered, and the solvent was removed in vacuo. Purification by silica gel chromatography (0-15% methanol in dichloromethane gradient) gave the desired compound, reference compound I-540 (3 mg, 13% yield) as a yellow film.1

1H-NMR (500 MHz, CDCla) 8 ppm 8.47 (d, 1 H), 8.27 (d, 1 H), 7.30 (s, 1 H), 7.24-7.20 (m , 1 H), 7.07-6.98 (m, 2 H), 6.91 (m, 1 H), 6.80 (br s, 1 H), 6.59 (d, 1 H), 6.10 (d, 1 H), 5.97 (d, 1 H), 5.92 (d, 1 H), 5.84 (quint, 1 H), 5.74 (br s, 1 H) .

Reference compound I-568

A solution of reference compound I-418 (1 equiv.), diphenyl phosphorazidate (1.5 equiv.) and triethylamine (1.5 equiv.) in toluene was heated at 50 °C for 15 h. The solution was cooled to 23 °C and treated with sodium methanolate (3 equiv., 0.5 N solution in methanol). After stirring at 23 °C for 1 h, saturated aqueous sodium bicarbonate solution was added and the resulting solution was stirred for an additional 1 h. The reaction was diluted with water and dichloromethane, and the aqueous layer was extracted with dichloromethane. The organics were dried over magnesium sulfate, filtered, and the solvent was removed in vacuo to give the crude product. Purification by silica gel chromatography (0-15% gradient (7:1 = acetonitrile:methanol) in dichloromethane) provided the desired compound, reference compound I-568 (11 mg, 52% yield) as a solid. white.

1H-NMR (500 MHz, CD3OD) 8 ppm 8.76 (d, 1 H), 8.21 (d, 1 H), 7.49 (s, 1 H), 7.29-7.24 (m , 1 H), 7.12-7.06 (m, 1 H), 7.03 (t, 1 H), 6.94 (d, 1 H), 6.80 (t, 1 H), 5 .97 (s, 2 H), 5.67-5.58 (m, 1 H), 3.73 (dd, 1 H), 3.51 (s, 3 H), 3.42 (dd, 1 H).

Reference compound I-576

The title compound was synthesized in 2 steps:

Step 1: Synthesis of (S)-1-((2-(1-(2-fluorobenzyl)-5-(isoxazol-3-yl)-1H-pyrazol-3-yl)pyrimidin-4-yl) acetate amino)-1-oxopropan-2-yl

The intermediate product was prepared following general procedure C, except that (S)-2-acetoxypropanoic acid was the acid reagent (3 equiv.), 10 equivalents of triethylamine, and 4 equivalents of T3P were used. The solution was heated to 50 °C for 16 h, at which time the solution was diluted with ethyl acetate and water. The layers were separated and the organic layer was washed with water and saturated aqueous sodium chloride. The organic substances were dried over magnesium sulfate, filtered, and the solvent was removed in vacuo. Purification by silica gel chromatography (0-5% methanol in dichloromethane gradient) provided the impure intermediate, which was taken up in the next step without further manipulation.

Stage 2: Synthesis of reference compound I-576

To a solution of (S)-1-((2-(1-(2-fluorobenzyl)-5-(isoxazol-3-yl)-1H-pyrazol-3-yl)—pyrimidin-4-yl) acetate amino)-1-oxopropan-2-yl (1 equiv.) in 4:1=methanol:water, potassium carbonate (0.5 equiv.) was added in one part. After stirring for 10 minutes, the reaction solution was acidified with 3N hydrochloric acid solution and diluted with water and dichloromethane. The layers were separated and the aqueous layer was extracted with dichloromethane. The organic substances were dried over magnesium sulfate, filtered, and the solvent was removed in vacuo. Purification by silica gel chromatography (0-5% methanol in dichloromethane) provided the desired compound, reference compound I-576 (4.5 mg, 29% yield) as a white solid.

1H-NMR (500 MHz, CD3OD) 8 ppm 8.77 (m, 1 H), 8.71 (d, 1 H), 8.14 (d, 1 H), 7.54 (s, 1 H) , 7.30-7.26 (m, 1 H), 7.11 7.03 (m, 2 H), 6.91-6.88 (m, 2 H), 5.98 (s, 2 H) ), 4.32 (q, 1 H), 1.45 (d, 3 H).

Reference compound I-580

The title compound was prepared following general procedure C, except that 2-methyl-2-(methylsulfonyl)propanoic acid was the acid reagent (3 equiv.), 10 equivalents of triethylamine were used, and 4 equivalents of the T3P. The solution was heated to 70 °C for 3 h, at which time the solution was diluted with ethyl acetate and water. The layers were separated and the organic layer was washed with water and saturated aqueous sodium chloride. The organic substances were dried over magnesium sulfate, filtered, and the solvent was removed in vacuo. Purification by silica gel chromatography (gradient 0-100% ethyl acetate in hexanes) gave the desired compound, reference compound I-580 (30 mg, 42% yield) as a yellow foam.

1H-NMR (500 MHz, CDCh) 89.13 (br s, 1 H), 8.78 (d, 1 H), 8.48 (d, 1 H), 8.06 (d, 1 H), 7.46 (s, 1 H), 7.24-7.19 (m, 1 H), 7.06-7.02 (m, 1 H), 7.00-6.97 (m, 1 H) ), 6.89-6.86 (m, 1 H), 6.63 (d, 1 H), 6.03 (s, 2 H), 3.00 (s, 3 H), 1.81 ( s, 6H).

Reference compound I-582

The title compound was prepared following general procedure C, except that 5-fluoro-2-(1-(2-fluorobenzyl)-5-(isoxazol-3-yl)-1H-pyrazol-3-yl)pyrimidin was used -4-amine (intermediate product described in a previous patent: WO2012/003405 A1; 1 equiv.) instead of intermediate 2, the intermediate 2-methyl-2-(methylsulfonyl)propanoic acid was the acid reagent (3 equiv. ), 10 equivalents of triethylamine were used, and 4 equivalents of propylphosphonic anhydride (T3P, 50% by weight in ethyl acetate) were used. After heating the mixture at 90 °C for 4 hours, the solution was diluted with ethyl acetate and water. The layers were separated and the aqueous layer was extracted with ethyl acetate. The organic substances were washed with water, dried over magnesium sulfate, filtered, and the solvent was removed in vacuo. Purification by silica gel chromatography (gradient (7:1 acetonitrile:methanol) in dichloromethane) provided the desired compound, reference compound I-582 (29 mg, 41% yield) as a white solid.

1H-NMR (500 MHz, CDCh) 89.10 (br s, 1 H), 8.67 (d, 1 H), 8.48 (d, 1 H), 7.42 (s, 1 H), 7.23-7.19 (m, 1 H), 7.06-7.02 (m, 1 H), 7.10-6.97 (t, 1 H), 6.86 (t, 1 H ), 6.63 (d, 1 H), 6.02 (s, 2 H), 3.02 (s, 3 H), 1.82 (s, 6 H).

Reference compound I-587

The composition of the title was prepared following the general management procedure C, except for the acid 2 - (methyl sulfonyl) propanic. It was the acidic reagent (3 eq u iv.), 10 eq u i v a l e n t e s of three t i la mine were used, and 4 eq u i v a l e n t e s of T 3 P were used. After stirring for 1 hour at 70°C, the reaction mixture was diluted with ethyl acetate and water. The layers were separated and the aqueous layer was extracted with ethyl acetate. The organic substances were washed with water, dried over magnesium sulfate, filtered, and the solvent was removed in vacuo. Purification by silica gel chromatography (0-5% methanol in dichloromethane gradient) provided the desired compound, reference compound I-587 (45 mg, 64% yield) as a brown solid.

1H-NMR (500 MHz, CDCh) 89.45 (br s, 1 H), 8.72 (d, 1 H), 8.48 (d, 1 H), 8.01 (d, 1 H), 7.34 (s, 1 H), 7.21-7.16 (m, 1 H), 7.04-7.01 (m, 1 H), 6.92 (t, 1 H), 6, 80-6.77 (m, 1 H), 6.65 (d, 1 H), 6.00 (s, 2 H), 4.14 (q, 1 H), 3.02 (s, 3 H ), 1.73 (d, 3H).

Reference compound I-609

To a solution of potassium ((2,2,2-trifluoroethyl)sulfonyl)amide (2 equiv.) in dimethyl sulfoxide was added intermediate 1 (1 equiv.). After stirring for 62 h, the solution was diluted with ethyl acetate and 1N aqueous hydrochloric acid solution. The layers were separated and the aqueous layer was extracted with ethyl acetate. The organic substances were washed with water and brine, dried over magnesium sulfate, filtered, and the solvent was removed in vacuo. The crude residue was taken up in methanol and the resulting solid was filtered and washed with additional methanol. The residue was taken up again in methanol and the resulting solids were filtered and rinsed with methanol to provide the desired compound, reference compound I 609 (24 mg, 28% yield) as a solid.

1H-NMR (500 MHz, DMSO-d6) 89.14 (d, 1 H), 8.44 (br s, 1 H), 7.51 (s, 1 H), 7.37-7.31 ( m, 1 H), 7.24-7.20 (m, 2 H), 7.12 (t, 1 H), 7.02-6.99 (m, 1 H), 5.92 (s, 2 H), 4.74 (br s, 2 H).

Reference compound I-627

A suspension of 2-(1-(2-fluorobenzyl)-5-(isoxazol-3-yl)-1H-pyrazol-3-yl)pyrimidin-4-ol (intermediate product described in WO2012/003405 A1; 1 equiv. ) in phosphoryl trichloride (20 equiv.) was heated at 60 °C for 2 h. After evaporating to dryness in vacuo, the resulting residue and 2-(aminomethyl)-3,3,3-trifluoro-2-hydroxypropanamide (3 equiv.) were dissolved in a 2:1 mixture of dioxane:water and treated with triethylamine (10 equiv.). The solution was heated at 110 °C for 38 h. The solution was diluted with dichloromethane and 1N hydrochloric acid, the layers were separated and the aqueous layer was extracted with dichloromethane. The organic substances were dried over magnesium sulfate, filtered, and the solvent was removed in vacuo. Purification by silica gel chromatography (0-65% gradient (7:1=acetonitrile:methanol) in dichloromethane) provided the contaminated product. The crude product was partitioned between water and dichloromethane. The layers were separated and the organic layer was washed with water. The organic layer was dried over magnesium sulfate, filtered, and the solvent was removed in vacuo to provide the desired compound, reference compound I-627 (1 mg, 1% yield) as a transparent film.

1H-NMR (500 MHz, CD3OD) 88.78 (s, 1 H), 8.17 (d, 1 H), 7.42 (s, 1 H), 7.30-7.26 (m, 1 H), 7.11-7.04 (m, 2 H), 7.01 6.98 (m, 1 H), 6.89 (s, 1 H), 6.59 (d, 1 H), 5.97 (s, 2 H), 4.10-4.02 (m, 2 H).

Reference compound I-634

The title compound was prepared following general procedure B, except that 3-amino-1,1,1-trifluoro-2-methylpropan-2-ol (as HCl salt, 2 equiv.) was the amine reagent, and 4 equivalents of triethylamine were used. After stirring at 90 °C for 21 h, preparation and purification by silica gel chromatography (0-70% ethyl acetate in hexanes gradient) afforded the desired compound, reference compound I-634 (33 mg , yield 48%) as a white solid.

1H-NMR (500 MHz, CDCla) 8 ppm 8.48 (m, 1 H), 8.22 (d, 1 H), 7.27 (s, 1 4-7.19 (m, 1 H), 7.05-6.95 (m, 3 H), 6.61 (m, 1 H), 6.04-5.93 (m, 3 H), 5.74 (br s, 1 H), 4 .00 (dd, 1 H), 3.81 (dd 1 H), 1.44 (s, 3 H).

Reference compound I-631

The title compound was prepared following general procedure B, except that 4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridin-3(2H)-one dihydrochloride (2 equiv) was the amine reagent, and 4 equivalents of triethylamine were used. After stirring at 90 °C for 1.5 hours, preparation and purification by silica gel chromatography (0-15% methanol in dichloromethane gradient) provided the desired compound, reference compound I-631 (9 mg, yield 14%) as a white solid.1

1H-NMR (500 MHz, CD3OD) 8 ppm 8.79 (d, 1 H), 8.22 (d, 1 H), 7.50 (s, 1 H), 7.31-7.27 (m , 1 H), 7.13-7.09 (m, 1 H), 7.05 (t, 1 H), 6.94 (m, 1 H), 6.85-6.82 (m, 1 H), 5.98 (s, 2 H), 4.76 (s, 2 H), 4.17 (t, 2 H), 2.85 (t, 2 H).

Reference compound I-328

The title compound was prepared following general procedure C, except that 1-(trifluoromethyl)cyclopropanecarboxylic acid (2 equiv.) was the acid reagent, 3 equivalents of triethylamine were used, and 3 equivalents of T3P were used. The solution was stirred at 23 °C for 18 h, at which time the solution was diluted with water and extracted with ethyl acetate. The organic phase was washed with water (2 times) and saturated aqueous sodium chloride. The organic substances were dried over sodium sulfate, filtered, and concentrated in vacuo. Purification of the residue by silica gel chromatography (ethyl acetate in hexanes) provided the desired compound, reference compound I-328 (2 mg, 14% yield).

1H-NMR (400 MHz, CDCh) 8 ppm 8.73 (d, 1 H), 8.57 (br s, 1 H), 8.45 (d, 1 H), 8.02 (d, 1 H ), 7.45 (s, 1 H), 7.19 (m, 1 H), 7.02 (m, 1 H), 6.96 (m, 1 H), 6.83 (m, 1 H ), 6.60 (d, 1 H), 6.01 (s, 2 H), 1.40 (m, 4 H).

Reference compound I-415

The title compound was prepared following general procedure C, except that 5-fluoro-2-(1-(2-fluorobenzyl)-5-(isoxazol-3-yl)-1H-pyrazol-3-yl)pyrimidin was used -4-amine (intermediate product described in a previous patent: WO2012/003405 A1; 1 equiv.) instead of intermediate 2, 1-(trifluoromethyl)cyclopropanecarboxylic acid (3 equiv.) was the acid reagent, 7 were used equivalents of triethylamine and 4 equivalents of T3P. After heating at 90 °C for 2 d, the vial was cooled to 23 °C and the contents were diluted with ethyl acetate. The contents were washed with water (3 times), brine, then dried with sodium sulfate, filtered and concentrated by rotary evaporation. Purification of the residue by silica gel chromatography (ethyl acetate in hexanes) provided the desired compound, reference compound I-415 (42 mg, 30% yield) as a white solid.

1H-NMR (400 MHz, CDCh) 8 ppm 8.67 (s, 1 H), 8.49 (s, 1 H), 8.31 (br s, 1 H), 7.43 (s, 1 H) ), 7.23 (m, 1 H), 7.06 (m, 1 H), 6.99 (m, 1 H), 6.84 (m, 1 H), 6.63 (s, 1 H ), 6.04 (s, 2 H), 1.64 (m, 2 H), 1.45 (m, 2 H).

Reference compound I-460

The title compound was prepared following general procedure C, except that 5-fluoro-2-(1-(2-fluorobenzyl)-5-(isoxazol-3-yl)-1H-pyrazol-3-yl)pyrimidin was used -4-amine (intermediate product described in a previous patent: WO2012/003405 A1; 1 equiv.) instead of intermediate 2, 1-cyanocyclopropanecarboxylic acid (3 equiv.) was the acid reagent, 7 equivalents of triethylamine were used and 4 T3P equivalents. After heating at 50 °C for 18 h, the vial was cooled to 23 °C and the contents were diluted with ethyl acetate. The contents were washed with water (3 times), brine, then dried with sodium sulfate, filtered and concentrated by rotary evaporation. Purification of the residue by silica gel chromatography (ethyl acetate in hexanes) provided the desired compound, reference compound I-460 (29 mg, 23% yield) as a white solid.

1H-NMR (400 MHz, CDCla) 8 ppm 8.61 (s, 1 H), 8.50 (br s, 1 H), 8.41 (s, 1 H), 7.34 (s, 1 H) ), 7.14 (m, 1 H), 6.97 (m, 1 H), 6.91 (m, 1 H), 6.77 (m, 1 H), 6.54 (s, 1 H ), 5.95 (s, 2 H), 1.80 (m, 2 H), 1.67 (m, 2 H).

Reference compound I-483

The title compound was synthesized in 2 steps:

Step 1: Synthesis of 3-(3-(4-chloro-5-nitropyrimidin-2-yl)-1-(2-fluorobenzyl)-1H-pyrazol-5-yl)isoxazole

2-(1-(2-fluorobenzyl)-5-(isoxazol-3-yl)-1H-pyrazol-3-yl)-5-nitropyrimidin-4-ol was added (intermediate product described in a previous patent: WO2012/ 003405 A1; 1 equiv.) to phosphorus oxychloride (22 equiv.) and the mixture was heated for 4 h at 90 °C. The content was concentrated in vacuo and the residue was taken up in ethyl acetate and subsequently washed with 10% sodium bicarbonate solution (2 times), water (2 times) and brine. The organic layer was dried over sodium sulfate, filtered and concentrated in vacuo to provide the desired intermediate, 3-(3-(4-chloro-5-nitropyrimidin-2-yl)-1-(2-fluorobenzyl)- 1 HOUR-

-pyrazol-5-yl)isoxazole as a tan solid (1.86 g, 96% yield).

1H-NMR (400 MHz, CDCh) 8 ppm 9.35 (s, 1 H), 8.53 (d, 1 H), 7.59 (s, 1 H), 7.25 (m, 1 H) , 7.07 (m, 1 H), 7.02 (m, 1 H), 6.91 (m, 1 H), 6.65 (d, 1 H), 6.08 (s, 2 H) .

Stage 2: Synthesis of reference compound I-483

The title compound was prepared following general procedure B, except that 3-(3-(4-chloro-5-nitropyrimidin-2-yl)-1-(2-fluorobenzyl)-1H-pyrazol-5-yl was used )isoxazole instead of intermediate 1, 2-(aminomethyl)-1,1,1,3,3,3-hexafluoropropan-2-ol (1.5 equiv.) was the amine reagent, 3 equivalents were used of triethylamine, and the content was heated at 30 °C for 1 h as a solution in dioxane:water (3:1). The reaction was cooled and diluted with ethyl acetate. The organic layer was washed with water (2 times) and brine, then dried with sodium sulfate, filtered and concentrated in vacuo. The crude material was purified by silica gel chromatography with a gradient of 0-100% ethyl acetate/hexanes providing the desired compound, reference compound I-483 (77 mg, yield 73%) as a white solid.

1H-NMR (400 MHz, CDCh) 8 ppm 9.36 (s, 1 H), 8.59 (m, 1 H), 8.55 (d, 1 H), 7.64 (br s, 1 H ), 7.42 (s, 1 H), 7.28 (m, 1 H), 7.08 (m, 1 H), 7.06 (m, 1 H), 6.64 (d, 1 H ), 5.98 (s, 2 H), 4.27, (d, 2 H).

REFERENCE COMPOSITION I - 484

A solution of the reference compound I-483 (1 equiv.) in methanol at 23 °C was treated with 10% palladium on carbon (0.2 equiv.), then placed under an atmosphere of H2 supplied by balloon filled with hydrogen attached to a needle. The mixture was stirred for 1 h under positive H2 pressure and filtered through Celite. The filter cake was rinsed with methanol, and the washed substances were concentrated in vacuo. The resulting residue was purified by silica gel chromatography using a gradient of ethyl acetate in hexanes to provide the desired compound, reference compound I-484 (53 mg, 66% yield) as a white solid.

1H-NMR (400 MHz, CDCh) 8 ppm 9.39 (s, 1H), 7.92 (br s, 1H), 7.19 (m, 1H), 7.13 (m, 2H), 7, 98 (m, 1H), 6.92 (m, 2H),

6.52 (s, 1H), 5.85 (s, 2H), 4.01, (s, 2H).

Reference compound I-541

A mixture of 3-(3-(4-chloro-5-nitropyrimidin-2-yl)-1-(2-fluorobenzyl)-1H--pyrazol-5-yl)isoxazole (as described in synthesis step 1 of reference compound I-483 (1 equiv.), methyl carbamate (5 equiv.) and cesium carbonate

(5 equiv.) was heated at 90 °C for 18 h. After cooling to 23 °C, the mixture was diluted with ethyl acetate and washed with water (3 times) and brine. The contents were dried over sodium sulfate, filtered, and concentrated in vacuo. The resulting residue was purified by silica gel chromatography using a gradient of ethyl acetate in hexanes to provide the desired compound, reference compound I-541 (35 mg, 19% yield) as a light yellow solid.

1H-NMR (400 MHz, CDCla) 8 ppm 10.09 (br s, 1 H), 9.51 (s, 1 H), 8.50 (d, 1 H), 7.60 (s, 1 H) ), 7.24 (m, 1 H), 7.06 (m, 1

H), 7.02 (m, 1 H), 6.87 (m, 1 H), 6.67 (d, 1 H), 6.07 (s, 2 H), 3.95 (s, 3 H).

Reference compound I-542

A solution of reference compound I-541 (1 equiv.) in methanol at 23 °C was treated with 10% palladium on carbon (0.2 equiv.), then placed under an atmosphere of H2 supplied by balloon filled with hydrogen attached to a needle. The mixture was stirred for 1 h under positive H2 pressure and filtered through Celite. The filter cake was rinsed with methanol, and the washed substances were concentrated in vacuo. The resulting residue was purified by silica gel chromatography using a gradient of ethyl acetate in hexanes to provide the desired compound, reference compound I-542 (26 mg, 87% yield) as a solid.

1H-NMR (400 MHz, CDCta) 8 ppm 8.43 (d, 1 H), 8.27 (s 1 H), 7.52 (s, 1 H), 7.32 (s, 1 H), 7.18 (m, 1 H), 7.02 (m, 1 H), 6.99 (m, 1 H), 6.77 (m, 1 H), 6.57 (d, 1 H), 5, 99 (s, 2 H), 4.61 (br s, 2 H), 3.81 (s, 3 H).

Reference compound I-488

The title compound was prepared following general procedure B, except that 2-(aminomethyl)-1,1,1,3,3,3-hexafluoropropan-2-ol (2.3 equiv.) was the amine reagent. , 3-(3-(4-chloro-5-fluoropyrimidin-2-yl)-1-(2,3-difluorobenzyl)-1H-pyrazol-5-yl)isoxazole (described in step 2 for the synthesis) was used of reference compound I-235) instead of intermediate 1 and the contents were heated at 90 °C for 3 days. The contents were cooled to 23°C, and the mixture was diluted with ethyl acetate. The organic layer was washed with water (2 times) and brine, then dried with sodium sulfate, filtered and concentrated in vacuo. The crude material was purified by silica gel chromatography (gradient of ethyl acetate in hexanes) providing the desired compound, reference compound I-488

(62 mg, 44% yield) as a white solid.

1H-NMR (400 MHz, CDCta) 8 ppm 8.52 (s, 1 H), 8.39 (s, 1 H), 8.29 (s, 1 H), 7.25 (s, 1 H) , 7.09 (m, 1 H), 6.98 (m, 1 H) 6.96 (m, 1 H), 6.64 (s, 1 H), 5.99 (s, 2 H), 5 .58 (br s, 1 H), 4.14 (d, 2 H).

Reference compound I-489

The title compound was prepared following general procedure B, except that 2-(aminomethyl)-3,3,3-trifluoro-2-hydroxypropanoic acid (1.3 equiv.) was the amine reagent, 3- (3-(4-chloro-5-fluoropyrimidin-2-yl)-1-(2,3-difluorobenzyl)-1H-pyrazol-5-yl)isoxazole (described in step 2 for the synthesis of reference compound I -235) instead of intermediate 1, and the contents were heated at 90 °C for 3 days. The contents were cooled to 23 °C, and the mixture was diluted with water, and the pH was adjusted to 5 with a 3N HCl solution. The mixture was filtered, and the filter cake was washed with water (2 times) and dried under vacuum. A part of the residue was dissolved in dichloromethane/methanol (4 ml, 1:1).

After filtration, 62 mg of insoluble material remained. The soluble fraction was subjected to silica gel chromatography using a dichloromethane/methanol gradient providing the reference compound I-489 as a white solid (60 mg). Analysis of the insoluble material remaining after filtration also showed to be the reference compound I-489 (total: 0.122 g, 90% yield). 1H-NMR (400 MHz, CD3OD) 8 ppm 8.77 (d, 1 H), 8.16 (d, 1 H), 7.51

(s, 1 H), 7.15 (m, 1 H), 7.01 (m, 1 H), 6.94 (d, 1 H), 6.71 (m, 1 H), 6.00 (s, 2 H), 4.43 (d, 1 H), 4.12 (d, 1 H).

Reference compound I-522

A solution of the compound of reference I - 489 (1 eq u iv .) in i chloro m e t a n o s is t ra t ed on the basis of H u n i g (3 eq u iv .) and

H A T U ( 1 , 5 eq u iv . ) . The final solution was stirred for 2 h, then a solution of ammonia (0.5 M in dioxane, 8 equiv.) was added to the reaction. The contents were allowed to stir overnight at 23°C. The mixture was diluted with dichloromethane and washed with water (3 times) and brine. The solution was then dried over sodium sulfate, filtered and concentrated in vacuo. Purification of the crude residue by silica gel chromatography using a dichloromethane/methanol gradient provided the desired compound, reference compound I-522 (5 mg, 8% yield) as a white solid.

1H-NMR (400 MHz, CDCh) 8 ppm 8.45 (s, 1 H), 8.14 (s, 1 H), 7.98 (s, 1 H), 7.18 (s, 1 H) , 7.13 (s, 1 H), 7.01 (m, 1 H), 6.90 (m, 2 H), 6.56 (s, 1 H), 5.90 (s, 2 H) , 5.65 (br s, 1 H), 5.53 (br s, 1 H), 4.12 (d, 2 H).

Reference compound I-507

The title compound was synthesized in 4 steps:

Stage 1: Synthesis of (3,3,3-trifluoropropyl)hydrazine hydrochloride

3-Bromo-1,1,1-trifluoropropane (1 equiv.) and hydrazine hydrate (10 equiv.) were dissolved in absolute ethanol and heated at 80 °C for 18 h. The solution was cooled to 23 °C and concentrated in vacuo at 15 °C. The thick oil was diluted with water and dichloromethane, then solid potassium carbonate was added to saturate the aqueous layer. The phases were mixed and separated, then the aqueous phase was extracted with additional dichloromethane (2 times). The combined organic phases were dried over sodium sulfate, filtered and concentrated in vacuo to provide a colorless oil. A small portion of the neutral hydrazine product was removed for characterization by NMR. The remainder was taken up in diethyl ether and treated with hydrochloric acid (2.5 M solution in ethanol), and the resulting mixture was concentrated in vacuo providing the desired intermediate product (3,3,3-trifluoropropyl)hydrazine (2.02 g, yield 43%) as a white solid.

1H-NMR (400 MHz, CDCla) 8 ppm 3.18 (br s, 4 H), 3.02 (m, 2 H), 2.36 (m, 2 H).

Step 2: Synthesis of ethyl 3-(isoxazol-3-yl)-1-(3,3,3-trifluoropropyl)--1H-pyrazole-5-carboxylate

A solution of (3,3,3-trifluoropropyl)hydrazine hydrochloride (1 equiv.) in a mixture of ethanol and water (9:1) at 23 °C was treated with potassium carbonate (0.6 equiv.) followed by of ethyl 4-(isoxazol-3-yl)-2-(methoxy(methyl)amino)-4-oxobut-2-enoate (2 equiv.) generated in step 1 of general procedure A, using 1-(isoxazole -3-yl)ethanone in step 1). The solution was stirred for 2 d at 23 °C, then 6N hydrochloric acid (1.5 equiv.) was added dropwise to the reaction. The solvents were removed in vacuo and the residue was taken up in ethyl acetate. The organic substances were washed with water (5 times), brine, dried over sodium sulfate, filtered, and concentrated in vacuo. The crude residue was purified by silica gel chromatography using gradient of ethyl acetate in dichloromethane providing the desired pyrazole ester, 3-(isoxazol-3-yl)-1-(3,3,3-trifluoropropyl)-1H Ethyl-pyrazole-5-carboxylate (1.34 g, 36% yield) as a light yellow solid.1

1H-NMR (400 MHz, CDCh) 8 ppm 8.55 (d, 1 H), 7.15 (s, 1 H), 6.63 (d, 1 H), 4.95 (m, 2 H) , 4.46 (q, 2 H), 2.85 (m, 2 H), 1.44 (t, 3 H).

Step 3: Synthesis of 5-(isoxazol-3-yl)-1-(3,3,3-trifluoropropyl)-1H-pyrazole-3-carboximidamide

The desired amidine intermediate was generated according to the procedure described in step 3 of general procedure A, except for the use of 3-(isoxazol-3-yl)-1-(3,3,3-trifluoropropyl)-1H- ethyl pyrazole-5-carboxylate as the starting ester and the mixture was heated for 4 h at 110 °C. The reaction mixture was cooled on ice, then methanol (14 equiv.) and aqueous hydrochloric acid (17 equiv.) were added successively for 5 min. This mixture was heated for 30 min at 80 °C, then cooled on ice and filtered. The filter cake was washed with toluene (2 times) and air dried to provide the crude amidine hydrochloride salt. This material was stirred in saturated aqueous sodium carbonate and extracted with ethyl acetate/isopropyl alcohol (5:1 mixture). The organic phase was washed with water and brine, dried over sodium sulfate, filtered and concentrated in vacuo to provide the desired neutral amidine, 5-(isoxazol-3-yl)-1-(3,3,3- trifluoropropyl)-1H-pyrazole-3-carboximidamide as a light yellow solid.

1H-NMR (400 MHz, CDCh) 8 ppm 8.45 (d, 1 H), 6.99 (s, 1 H), 6.55 (d, 1 H), 5.61 (br. s., 3 H), 4.83-4.74 (m, 2 H), 2.81 2.65 (m, 2 H).

Stage 4: Synthesis of reference compound I-507

The title product was prepared following step 4 of general procedure A, except that 5-(isoxazol-3-yl)-1-(3,3,3-trifluoropropyl)-1H-pyrazole-3-carboximidamide was the amidine As a starting point, 2.5 equivalents of sodium (Z)-3-ethoxy-2-fluoro-3-oxoprop-1-en-1-olate were used and the mixture was heated for 2 h at 90 °C. The reaction was cooled to 23 °C, and the solvent was removed in vacuo. The residue was redissolved in dichloromethane and treated with hydrochloric acid (2.5 M in ethanol, 3 equiv.). The resulting solids were filtered, washed with dichloromethane (2 times) and air dried to provide the desired compound, reference compound I-507 (0.43 g, 110% yield) as a white solid.

1H-NMR (400 MHz, CD3OD) 8 ppm 8.84 (d, 1 H), 8.03 (d, 1 H), 7.40 (s, 1 H), 6.95 (d, 1 H) , 4.96 (t, 2 H), 2.92 (m, 2 H).

Reference compound I-511

The title compound was synthesized in 2 steps:

Step 1: Synthesis of 3-(3-(4-chloro-5-fluoropyrimidin-2-yl)-1-(3,3,3-trifluoropropyl)-1H-pyrazol-5-yl)isoxazole

A mixture of reference compound I-507 (1 equiv.) in phosphorus oxychloride (28 equiv.) was heated for 2 h at 90 °C. The solvent was removed in vacuo and the residue was rinsed with dichloromethane (2 times) providing the desired intermediate product 3-(3-(4-chloro-5-fluoropyrimidin-2-yl)-1-(3,3,3- trifluoropropyl)-1H-pyrazol-5-yl)isoxazole (0.28 g, 69% yield) as a tan solid.

1H-NMR (400 MHz, CDCla) 8 ppm 8.59 (s, 1 H), 8.47 (d, 1 H), 7.30 (s, 1 H), 6.60 (d, 1 H) , 4.92 (t, 2 H), 2.81 (m, 2 H).

Stage 2: Synthesis of reference compound I-511

The title compound was prepared following general procedure B, except that 2-aminoacetamide (as HCl salt, 2 equiv.) was the amine reagent, 3-(3-(4-chloro-5-fluoropyrimidin- 2-yl)-1-(3,3,3-trifluoropropyl)-1H-pyrazol-5-yl)isoxazole (described above) instead of intermediate 1, 5 equivalents of triethylamine were used, and the contents were heated to 90 °C for 2 h. The mixture was cooled to 23 °C, diluted with water and the pH was collected at ~5 with aqueous 3N hydrochloric acid. The crude product was collected by filtration, then purified by silica gel chromatography with a gradient of methanol in dichloromethane providing the desired compound, reference compound I-511 (12 mg, 45% yield) as a white solid.

1H-NMR (400 MHz, CD3OD) 8 ppm 8.81 (s, 1 H), 8.13 (d, 1 H), 7.34 (s, 1 H), 6.93 (s, 1 H) , 4.91 (m, 2 H), 4.21 (s, 2 H), 2.89 (m, 2 H).

The reference compound I-513

3-(3-(4-chloro-5-fluoropyrimidin-2-yl)-1-(3,3,3-trifluoropropyl)-1H-pyrazol-5-yl)isoxazole (described in step 1 of synthesis of the compound Reference I-511 (1 equiv.) and concentrated aqueous ammonium hydroxide (2.8 equiv.) in dioxane were sealed in a screw cap vial and heated for 2 h at 95 °C. The mixture was cooled to 23. °C, diluted with water, then filtered. The filter cake was washed with water (2 times) and air dried to provide the desired compound, reference compound I-513 (0.14 g, yield 76). %) as a light tan powder.1*3

1H-NMR (400 MHz, CDCta) 8 ppm 8.51 (d, 1 H), 8.21 (d, 1 H), 7.26 (s, 1 H), 6.63 (d, 1 H) , 5.26 (s, 2 H), 4.95 (m, 2 H), 2.85 (m, 2 H).

Reference compound I-516 and reference compound I-517

The title compound was prepared following general procedure B, except that 2-(aminomethyl)-1,1,1,3,3,3-hexafluoropropan-2-ol (2 equiv.) was the amine reagent, used 3-(3-(4-chloro-5-fluoropyrimidin-2-yl)-1-(3,3,3-trifluoropropyl)-1H-pyrazol-5-yl)isoxazole (described in step 1 of the synthesis of reference compound I-511 (1 equiv.) instead of intermediate 1, 6 equivalents of triethylamine were used, and the content was heated at 95 °C for 3 d. The mixture was cooled to 23 °C, diluted. with ethyl acetate, then washed with water (2 times) and brine. The contents were dried with sodium sulfate, filtered and concentrated in vacuo to give the crude product. The residue was purified by silica gel chromatography using a. hexane/ethyl acetate gradient providing two products, reference compound I-516 (27 mg, 37% yield) as a white solid, and reference compound I-517 (9 mg, 16% yield) as a solid white.

1H-NMR for Reference Compound I-517 (400 MHz, CDCI3) 5 ppm 8.54 (s, 1 H), 8.15 (d, 1 H), 7.17 (s, 1 H), 6 .68 (s, 1 H), 4.94 (m, 2 H), 3.69 (q, 4 H), 2.89 (m, 2 H), 1.31 (t, 6 H).

Reference compound I-523

The title compound was prepared following general procedure B, except that 1-((methylamino)methyl)cyclopropanecarboxylic acid (as the HCl salt) was the amine reagent, 3-(3-(4-chloro- 5-fluoropyrimidin-2-yl)-1-(3,3,3-trifluoropropyl)-1H-pyrazol-5-yl)isoxazole (described in step 1 of synthesis of reference compound I-511 (1 equiv.) Instead of intermediate 1, 6.6 equivalents of triethylamine were used and the contents were heated at 90 °C for 18 h. The reaction mixture was diluted with water and carefully brought to pH 4 with an aqueous hydrochloric acid solution. 3N. The mixture was filtered, and the filter cake was washed with water (2 times) and air dried to provide the desired compound, reference compound I-523 (9 mg, 60% yield) as a white solid.

1H-NMR (400 MHz, CDCla) 5 ppm 8.51 (d, 1 H), 8.16 (d, 1 H), 7.16 (s, 1 H), 6.64 (s, 1 H) , 4.92 (m, 2 H), 4.10 (s, 2 H), 3.35 (d, 3 H), 2.87 (m, 2 H), 1.44 (m, 2 H) , 1.13 (m, 2 H).

Reference compound I-573

The title compound was synthesized in 2 steps:

Stage 1: Synthesis of 3-(methylsulfonyl)propanoic acid

A solution of 3-(methylthio)propanoic acid (1 equiv.) in glacial acetic acid was cooled on ice as aqueous hydrogen peroxide (27%, 6 equiv.) at a rate to maintain the internal temperature <50 °C. The cooling bath was removed and stirring was continued for 18 h at 23 °C. The solvents were removed in vacuo providing a white paste. The paste was mixed in dichloromethane and filtered. The filter cake was washed with additional dichloromethane (3 times) and air dried to provide the desired carboxylic acid, 3-(methylsulfonyl)propanoic acid (3.0 g, 47% yield) as a white solid. 1H-NMR (400 MHz, acetone-d6) 5 ppm 10.2 (br s, 1 H), 3.38 (t, 2 H), 3.00 (s, 3 H), 2.85 (t, 2H).

The title compound was prepared following general procedure C, except that 3-(methylsulfonyl)propanoic acid (2 equiv.) was the amine reagent, 6 equivalents of triethylamine and 3 equivalents of propylphosphonic anhydride (T3P, 50%) were used. by weight in ethyl acetate), and the solution was heated at 70 °C for 18 h. The contents were cooled to 23 °C, diluted with ethyl acetate, then washed with water (3 times) and brine. The contents were dried over sodium sulfate, filtered, and concentrated in vacuo. Purification of the residue by silica gel chromatography using a gradient of ethyl acetate in hexanes provided the desired compound, reference compound I-573 (6 mg, 8% yield) as a white solid.

1H-NMR (400 MHz, ace tone^) 5 ppm 8.77 (d, 1 H), 8.58 (d, 1 H), 7.92 (d, 1 H), 7.40 (s, 1 H), 7.20 (m, 1 H), 7.03 (m, 1 H), 6.98 (m, 1 H), 6.94 (d, 1 H), 6.83 (m, 1 H), 5.88 (s, 2 H), 3.38 (t, 2 H), 3.07 (t, 2 H), 2.88 (s, 3 H).

Reference compound I-588

The title compound was synthesized in 6 steps:

Step 1: Synthesis of ethyl 3-((ferf-butoxycarbonyl)amino)-4,4,4-trifluorobutanoate

A solution of ethyl 3-amino-4,4,4-trifluorobutanoate hydrochloride (1 equiv.) in THF was cooled on ice as triethylamine (2.2 equiv.) was added over 5 min. Di-tert-butyl dicarbonate (2 equiv.) in THF was added to the vessel and the mixture was stirred for 2 d at 23 °C. The solvent was removed in vacuo and the residue was taken up in ethyl acetate, then washed successively with water, 10% aqueous sodium bicarbonate solution, water (3 times) and then brine. The contents were dried with sodium sulfate, filtered and concentrated in vacuo. Purification of the residue by silica gel chromatography using a gradient of ethyl acetate in hexanes provided the desired Boc-protected amine, ethyl 3-((ferf-butoxycarbonyl)amino)-4,4,4-trifluorobutanoate (1 .9 g, yield 74%) as a white solid.

Step 2: Synthesis of ferf-butyl (1,1,1-trifluoro-4-hydroxybutan-2-yl)carbamate

A solution of ethyl 3-((ferf-butoxycarbonyl)amino)-4,4,4-trifluorobutanoate (1 equiv.) in THF was cooled on ice as lithium aluminum hydride (2 M in THF, 2.5 equiv. .) for 5 min. The solution was stirred for 3 h at 23 °C, then cooled on ice and treated sequentially with water, 15% aqueous NaOH, and water. Stirring continued for 15 min at 23 °C, then the mixture was filtered through Celite and the filter cake was rinsed with ethyl acetate (4 times). The combined organic filtrates were dried over sodium sulfate, filtered, and concentrated in vacuo to provide the desired alcohol intermediate, ferf-butyl (1,1,1-trifluoro-4-hydroxybutan-2-yl)carbamate ( 0.26 g, yield 98%) as a white solid.

1H-NMR (400 MHz, CDCla) 5 ppm 4.79 (br d, 1 H), 4.44 (m, 1 H), 3.76 (m, 1 H), 3.68 (m, 1 H ), 2.63 (br s, 1 H), 2.08 (m, 1 H), 1.55 (m, 1 H), 1.45 (s, 9 H).

Step 3: Synthesis of 3-((ferf-butoxycarbonyl)amino)-4,4,4-trifluorobutyl methanesulfonate

A solution of ferf-butyl (1,1,1-trifluoro-4-hydroxybutan-2-yl)carbamate (1 equiv.) and triethylamine (2.5 equiv.) in dichloromethane was stirred at 23 °C as added methanesulfonyl chloride (1.7 equiv.) to the container. The resulting solution was stirred for 2 h at 23 °C, then diluted with ethyl acetate and washed with water (4 times) and brine. The contents were dried over sodium sulfate, filtered and concentrated in vacuo to provide the desired mesyl-protected alcohol intermediate, 3-((ferf-butoxycarbonyl)amino)-4,4,4-trifluorobutyl methanesulfonate (0 .29 g, yield 89%) as a colorless solid.

1H-NMR (400 MHz, CDCla) 5 ppm 4.67 (br d, 1 H), 4.45 (m, 1 H), 4.31 (m, 2 H), 3.05 (s, 3 H) ), 2.28 (m, 1 H), 1.84 (m, 1 H), 1.45 (s, 9 H).

Step 4: Synthesis of ferf-butyl (1,1,1-trifluoro-4-(methylsulfonyl)butan-2-yl)carbamate

A solution of 3-((ferf-butoxycarbonyl)amino)-4,4,4-trifluorobutyl methanesulfonate (1 equiv.) in THF at 23 °C was treated with sodium methanethiolate (10 equiv.), and the resulting solution It was heated for 6 h at 60 °C. The reaction mixture was then cooled on ice as m-chloroperoxybenzoic acid (70% w/w, 12.5 equiv.) was added in portions. To confirm complete conversion to sulfone, the reaction was analyzed by LC/Ms. The mixture was diluted with ethyl acetate and washed successively with 7:1=10% aqueous bicarbonate/3N aqueous sodium hydroxide solution (2 times), 10% aqueous sodium bicarbonate solution (2 times), water ( 4 times) and then brine. The contents were dried over sodium sulfate, filtered, and concentrated in vacuo to give a residue. The crude product was purified by silica gel chromatography, using a gradient of ethyl acetate in hexanes providing the desired sulfone intermediate, (1,1,1-trifluoro-4-(methylsulfonyl)butan-2-yl) ferf-butyl carbamate (0.18 g, 72% yield) as a white solid.1

1H-NMR (400 MHz, CDCta) 5 ppm 4.75 (br d, 1 H), 4.36 (m, 1 H), 3.16 (m, 2 H), 2.99 (s, 3 H) ), 2.37 (m, 1 H), 2.07 (m, 1 H), 1.49 (s, 9 H).

Step 5: Synthesis of 1,1,1-trifluoro-4-(methylsulfonyl)butan-2-amine

S e d is o l v i o ( 1 , 1 , 1 - tri f l u o r o - 4 - ( m e t i l su l f o n i l ) b u t a n - 2 - i l ) c a r b a m a t o d e f e r f - b u t i l 23 ° C, then treated with tri-fluous acid tic (25 eq u iv.) and was stirred for 3 h at 23 ° C. The reaction was diluted with 10% sodium bicarbonate aqueous solution, and the phases were c l a r e n well. The aqueous phase is extracted with methane (3 times), and the combined organic phases are dried over sulfate. sodium, were filtered, and concentrated in the vacuum itself by providing the intermediate amine protection product. , the 1 , 1 , 1 - tri fl u o ro - 4 - (m e t i l su l f o n i l ) b u t a n - 2-amine (18 mg, yield 54%) as a colorless oil.

1H-NMR (400 MHz, CDCh) 8 ppm 3.25 (m, 2 H), 3.15 (m, 1 H), 2.89 (s, 3 H), 2.23 (m, 1 H) , 1.80 (m, 1 H), 1.36 (br s, 2 H).

Stage 6: Synthesis of reference compound I-588

The title compound was prepared following general procedure B, except that 1,1,1-trifluoro-4-(methylsulfonyl)butan-2-amine (6 equiv.) was the amine reagent, 6 equivalents of triethylamine were used , and the content was microwaved at 215 °C for 2 h as a solution in NMP. Direct purification of the resulting mixture by silica gel chromatography using a gradient of ethyl acetate in hexanes provided impure product. Further purification by reverse phase HPLC provided the desired compound, reference compound I-588 (17 mg, 16% yield).

1H-NMR (400 MHz, CDCla) 8 ppm 8.40 (d, 1 H), 8.23 (d, 1 H), 7.29 (s, 1 H), 7.13 (m, 1 H) , 6.97 (m, 1 H), 6.91 (m, 1 H), 6.81 (m, 1 H), 6.56 (d, 1 H), 5.93 (d, 1 H) , 5.88 (d, 1 H), 5.35 (m, 1 H), 5.18, (br s, 1 H), 3.19 (m, 1 H), 3.11 (m, 1 H), 2.86 (s, 3 H), 2.53 (m, 1 H), 2.21 (m, 1 H).

Reference compound I-626

The example compound was prepared following general procedure B, except that 1,1,1-trifluoro-4-(methylsulfonyl)butan-2-amine (1.3 equiv.) was the amine reagent, 3- (3-(4-chloro-5-fluoropyrimidin-2-yl)-1-(3,3,3-trifluoropropyl)-1H-pyrazol-5-yl)isoxazole (described in step 1 of synthesis of the reference compound I-511 (1 equiv.) instead of intermediate 1, 1.3 equivalents of triethylamine were used and the content was heated by microwave at 215 °C for 2.5 h as a solution in NMP. Resulting mixture by silica gel chromatography using a gradient of ethyl acetate in hexanes gave impure product. Further purification by reverse phase HPLC gave the desired compound, reference compound I-626 (1 mg, 1% yield).

1H-NMR (400 MHz, CDCh) 8 ppm 8.53 (d, 1 H), 8.32 (d, 1 H), 7.30 (s, 1 H), 6.71 (d, 1 H) , 5.44 (m, 1 H), 5.27 (br d, 1 H), 4.94 (m, 2 H), 3.21 (m, 2 H), 2.95 (s, 3 H ), 2.87 (m, 2 H), 2.61 (m, 1 H), 2.28 (m, 1 H).

Reference compound I-617

The title compound was prepared in 3 steps:

Step 1: Synthesis of 2-(2-(1-(2-fluorobenzyl)-5-(isoxazol-3-yl)-1H-pyrazol-3-yl)-5-nitropyrimidin-4-yl)-2-methylmalonate of diethyl.

A mixture of 3-(3-(4-chloro-5-nitropyrimidin-2-yl)-1-(2-fluorobenzyl)-1H-pyrazole-

- 5-yl)isoxazole (described in step 1 for the synthesis of reference compound I-483, 1.2 equiv.), ethyl 2-methylmalonate (1 equiv.) and potassium f-butoxide (0.9 equiv.) in THF was stirred at room temperature for 15 min. The solution was diluted with saturated aqueous ammonium chloride solution and ethyl acetate. The phases were separated and the aqueous phase was extracted twice with ethyl acetate. The combined organic phase was washed with brine, dried over anhydrous sodium sulfate, filtered and concentrated. The crude material was purified by silica gel chromatography (10-50% ethyl acetate in hexane gradient) providing the desired intermediate product 2-(2-(1-(2-fluorobenzyl)-5-(isoxazole-3- Ethyl yl)-1H-pyrazol-3-yl)-5--nitropyrimidin-4-yl)-2-methylmalonate (94.5 mg, 40% yield) as a yellow solid.

1H NMR (500 MHz, DMSO-d6) 8 ppm 9.56 (s, 1 H), 9.14 (d, 1 H), 7.69 (s, 1 H), 7.36 (s, 1 H) ), 7.30 (d, 1 H), 7.19-7.25 (m, 1 H), 7.12 (t, 1 H), 6.97-7.03 (m, 1 H), 5.95-5.97 (m, 2 H), 4.12-4.19 (m, 4 H), 1.94 (s, 3 H), 1.11 (t, 6 H).

Step 2: Synthesis of 2-(5-amino-2-(1-(2-fluorobenzyl)-5-(isoxazol-3-yl)-1H-pyrazol-3-yl)pyrimidin-4-yl)-2- diethyl methylmalonate.

A mixture of 2-(2-(1-(2-fluorobenzyl)-5-(isoxazol-3-yl)-1H-pyrazol-3-yl)--5-nitropyrimidin-4-yl)-2-methylmalonate diethyl (1 equiv.) and 20% palladium on carbon (0.5 equiv.) in ethanol and ethyl acetate (1:1) were stirred in a hydrogen atmosphere at 23 °C for 18 h. The reaction mixture was then filtered through celite and the residue was washed with ethyl acetate. The filtrate was concentrated in vacuo and the residue was collected in the next step without any further purification or characterization.

Stage 3: Synthesis of reference compound I-617

A solution of 2 - ( 5 - a m i n o - 2 - ( 1 - ( 2 - f l u o ro b e n c i l ) - 5 - ( i s o x a z o l - 3 - i l ) - 1 H - p i r a z o l - 3 - i l ) - p i r i m i d i n - 4 - i l ) - 2 - m e t i l m a l o n a t o d e t i l o in n e t a n o l and T H F (2: 1) was heated at 85 ° C for 16 h. The r e s u l t a n t solution is concentrated on the void and the residue is purified using reverse-phase HPLC 60% aceton ytrilo in water, with TFA of 1%)

Step 1: Synthesis of diethyl 2-(dicyanomethyl)-2-methylmalonate

A mixture of diethyl 2-bromo-2-methylmalonate (1 equiv.), malononitrile (1 equiv.) and potassium f-butoxide (1 equiv.) in THF was heated at reflux for 15 h. The mixture was diluted with ethyl acetate and saturated aqueous ammonium chloride solution and the phases were separated. The aqueous phase was extracted twice with ethyl acetate. The combined organic phase was washed with brine, dried over anhydrous sodium sulfate, filtered and concentrated in vacuo to give an oil. The oil was purified by silica gel chromatography, using a gradient of 10-15% ethyl acetate in hexane providing the desired intermediate, diethyl 2-(dicyanomethyl)-2-methylmalonate (5.76 g, yield 32 %) as a colorless oil.

1H NMR (500 MHz, CDCla) 8 ppm 4.53 (s, 1 H), 4 .27-4.39 (m, 4 H), 1.81 (s, 3 H), 1.33 (t, 6H).

Stage 2: Synthesis of reference compound I-618

A mixture of 5-(isoxazol-3-yl)-1-(3,3,3-trifluoropropyl)-1H-pyrazole-3-carboximidamide (generated in step 3 for the synthesis of reference compound I-507 (1 equiv.), diethyl 2-(dicyanomethyl)-2-methylmalonate (1.15 equiv.) and potassium bicarbonate (2 equiv.) in f-BuOH was heated at reflux for 5 h. The resulting solution was then concentrated. in vacuo and the residue was purified by silica gel chromatography using gradient of 0-5% methanol in dichloromethane providing the desired compound, reference compound I-618 (88.5 mg, 51% yield) as a white solid.

1H NMR (500 MHz, DMSO-d6) 8 ppm 11.31 (s, 1 H), 9.14 (d, 1 H), 7.34 (s, 1 H), 7.24 (d, 1 H ), 6.57 - 6.71 (m, 2 H), 4.85 (t, 2 H), 4.11 (t, 2 H), 2.85-2.98 (m, 2 H), 1.61 (s, 3 H), 1.12 (t, 3 H).

Reference compound I-619

Ammonia (7.0 M in MeOH, 200 equiv.) was added to the reference compound I-618 (1 equiv.). The reaction mixture was heated at 50 °C for 16 h. The resulting solution was then concentrated in vacuo and the residue was purified by reverse phase HPLC (gradient of 20-40% acetonitrile in water with 1% TFA) to provide the desired compound, reference compound I-619 (4, 7 mg, yield 31%) as a white solid.

1H NMR (500 MHz, DMSO-d6) 8 ppm 11.30 (s, 1 H), 9.14 (d, 1 H), 7.42 - 7.50 (m, 1 H), 7.36 ( s, 1 H), 7.25 (d, 1 H), 7 .17-7.22 (m, 1 H), 6 .67-6.92 (m, 2 H), 4 .83-4, 89 (m, 2 H), 2.86-2.99 (m, 2 H), 1.56 (s, 3 H).

Reference compound I-620

The title compound was prepared in 3 steps:

Step 1: Synthesis of (E)-2-(1-(2-fluorobenzyl)-5-(isoxazol-3-yl)-1H-pyrazol-3-yl)-5-(phenyldiazenyl)pyrimidin-4,6- diamine.

A mixture of 1-(2-fluorobenzyl)-5-(isoxazol-3-yl)-1H-pyrazole-3-carboximidamide (generated in step 3 for the synthesis of reference compound I-507 (1 equiv), ( E)-2-(phenyldiazenyl)malononitrile (1.2 equiv.) and potassium bicarbonate (2 equiv.) in f-BuOH was heated at reflux for 18 h. After cooling, the reaction mixture was concentrated in vacuo and. It was carried to the next step without further purification.

Stage 2: Synthesis of 2-(1-(2-fluorobenzyl)-5-(isoxazol-3-yl)-1H-pyrazol-3-yl)--pyrimidin-4,5,6-triamine.

A mixture of (E)-2-(1-(2-fluorobenzyl)-5-(isoxazol-3-yl)-1H-pyrazol-3-yl)--5-(phenyldiazenyl)pyrimidin-4,6-diamine (1 equiv.) and 20% palladium on carbon (0.5 equiv.) in DMF was stirred under a hydrogen atmosphere at 23 °C for 18 h. The reaction mixture was then filtered through celite and the residue was washed with DMF followed by a small part of methanol. The filtrate was concentrated in vacuo and the residue was suspended in ethyl acetate and a drop of methanol and stirred vigorously. The precipitate was filtered, washed with ethyl acetate and dried under vacuum to provide the desired triaminopyrimidine intermediate, 2-(1-(2-fluorobenzyl)-5-(isoxazol-3-yl)-1H-pyrazole-3. -yl)-pyrimidin-4,5,6-triamine (278 mg, 46% yield in 2 steps) as a dark yellow solid.

Stage 3: Synthesis of reference compound I-620

A solution of 2-(1-(2-fluorobenzyl)-5-(isoxazol-3-yl)-1H-pyrazol-3-yl)pyrimidin-4,5,6-triamine (1 equiv.) in pyridine at 0 °C was treated with methyl chloroformate (1 equiv.). The reaction mixture was slowly warmed to 23 °C and stirred for 18 h. The volatile compounds were removed in vacuo and the residue was dissolved in ethyl acetate and washed with water. The contents were dried over anhydrous sodium sulfate, filtered and concentrated to give a dark yellow solid. The crude material was purified by reverse phase HPLC (20-40% acetonitrile/water gradient with 1% TFA) to provide the desired compound, reference compound I-620 (15 mg, 26% yield) as a yellow solid. pale.

1H NMR (500 MHz, CD3OD) 8 ppm 8.82 (d, 1 H), 7.51 (s, 1 H), 7.27 - 7.35 (m, 1 H), 7.04 - 7, 15 (m, 2 H), 6.92 - 6.99 (m, 1 H), 6.89 (d, 1 H), 6.00 (s, 2 H), 3.78 (br. s. , 3 H), 3.34 - 3.35 (m, 1 H).

Reference compound I-621

A solution of reference compound I-620 (1 equiv.) and LiHMDS (1 M in toluene, 6 equiv.) was formed in THF at 0 °C and stirred for 20 min. Iodomethane (12 equiv.) was added to the reaction vessel, the mixture was heated to 23 °C and stirred for 1 h. The mixture was diluted with dichloromethane and saturated aqueous ammonium chloride solution, and the phases were separated. The aqueous phase was extracted twice with dichloromethane. The combined organic phases were dried over anhydrous magnesium sulfate, filtered, and concentrated. The crude material was purified by reverse phase HPLC (20-40% acetonitrile/water gradient with 1% TFA) providing the desired compound, reference compound I-621 (1.6 mg, 17% yield) as a yellow solid.

1H NMR (500 MHz, CD3OD) 8 ppm 8.83 (d, 1 H), 7.53 (s, 1 H), 7.27 - 7.35 (m, 1 H), 7.05 - 7, 15 (m, 2 H), 6.96 (t, 1 H), 6.90 (d, 1 H), 6.00 (s, 2 H), 3.66 - 3.86 (m, 3 H ), 3.13 (d, 3H).

Reference compound I-623

The title compound was prepared in 2 steps:

Stage 1: Synthesis of 4-amino-2-(1-(2-fluorobenzyl)-5-(isoxazol-3-yl)-1H-pyrazol-3-yl)--5-methyl-6-oxo-6, 7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-5-carbohydrazide.

A mixture of reference compound I-420 (1 equiv.), anhydrous hydrazine (325 equiv.), and water (11.2 equiv.) in methanol was heated at 50 °C for 2 h. The resulting solution was concentrated in vacuo. Excess hydrazine was azeotropically removed by treatment with methanol and dichloromethane providing the desired acylhydrazine intermediate, 4-amino-2-(1-(2-fluorobenzyl)-5-(isoxazol-3-yl)-1H-pyrazole. -3-yl)-5-methyl-6-oxo-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-5-carbohydrazide as a yellow solid. The material was used as is in the next step without further purification.

Stage 2: Synthesis of reference compound I-623

A mixture of 4-amino-2-(1-(2-fluorobenzyl)-5-(isoxazol-3-yl)-1H-pyrazol-3-yl)-5-methyl-6-oxo-6,7-dihydro -5H-pyrrolo[2,3-d]pyrimidin-5-carbohydrazide (1 equiv.) and A/-acetylimidazole (4 equiv.) in THF was stirred at 23 °C for 16 h. The resulting solution was concentrated in vacuo and the residue was purified by reverse phase HPLC (gradient of 20-80% acetonitrile in water, with 1% TFA) to provide the desired compound, reference compound I-623 (71 mg, yield 46%) as a white solid.

1H NMR (500 MHz, CD3OD) 8 ppm 8.80 (d, 1 H), 7.52 (s, 1 H), 7.25 - 7.32 (m, 1 H), 7.06 - 7, 14 (m, 1 H), 7.01 - 7.06 (m, 1 H), 6.87 - 6.92 (m, 2 H), 5.98 (s, 2 H), 2.66 ( s, 1 H), 2.00 (s, 3 H), 1.82 (s, 3 H).

Reference compound I-478

To a stirred solution of the reference compound I-461 (1 equiv.) in DMF and ethanol (3:2) was added 10% palladium on carbon (10 equiv.), and the reaction vessel was placed under an atmosphere of hydrogen using a balloon and needle. The contents were stirred for 18 h at 23 °C, and the mixture was filtered and concentrated in vacuo. The crude material was purified by reverse phase HPLC providing the desired compound, reference compound I-461 (5 mg, 19% yield) as a solid.

1H-NMR (500 MHz, DMSO-d6) 8 ppm 9.69 (s, 1 H), 9.11 (d, 1 H), 8.78 (d, 1 H), 8.00 (d, 1 H), 7.72 (s, 1 H), 7.31 - 7.38 (m, 1 H), 7.29 (d, 1 H), 7.21 - 7.26 (m, 1 H) , 7.12 (t, 1 H), 6.86-6.94 (m, 2 H), 5.95 (s, 2 H), 1.22- 1.28 (m, 2 H), 1 .09 - 1.14 (m, 2H).

Reference compound I-479

An agitated solution of the intermediate product 2 (1 eq u iv . ) and chloride 2 - m e t o x i e t a n o s u lf o n i l o (1 e q u iv . ) in n d i c l o r m e D B U (1 eq u iv.) was added. The reaction was heated to 60 ° C for 24 hours, then cooled to 23 ° C. The content was diluted in water and extracted with dichloromethane (3 times). The combined organic layers were washed with 1N HCl solution, dried over sodium sulfate, filtered, and concentrated in vacuo. The crude material was purified by reverse phase HPLC providing the desired compound, reference compound I-479 (8 mg, 6% yield) as a solid.

1H-NMR (500 MHz, CD3OD) 8 ppm 8.82 (d, 1 H), 8.50 (d, 1 H), 7.60 (s, 1 H), 7.27 - 7.35 (m , 1 H), 7.05 - 7.15 (m, 3 H), 6.94 - 6.98 (m, 2 H), 6.02 (s, 2 H), 3.86 (s, 4 H), 3.27 (s, 3 H).

Reference compound I-595

A solution of 2-(1-(2-fluorobenzyl)-5-(isoxazol-3-yl)-1H-pyrazol-3-yl)pyrimidin-5-amine (this compound was described in the previous patent application WO2012003405 A1 ) in dichloromethane/pyridine (2:1) was treated with 3,3,3 trifluoropropane-1-sulfonyl chloride (1.8 equiv.). After 3 h, 1N NaOH solution was added and the reaction was stirred for 1.5 h. Water was then added and the resulting mixture was acidified to pH 3 with 1N HCl solution and extracted with dichloromethane. The combined organic phases were dried over sodium sulfate, filtered, and the solvent was removed in vacuo. The crude material was purified by silica gel chromatography with a gradient of 0-10% methanol/dichloromethane providing the desired compound, reference compound I-595 (6.8 mg, 15% yield) as a white solid.

1H-NMR (500 MHz, CD3OD) 8 ppm 8.77 (d, 1 H), 8.74 (s, 2 H), 7.52 (s, 1 H), 7.27 (ap. q, 1 H), 7.09 (m, 1 H), 7.04 (ap. t, 1 H), 6.90 (d, 1 H), 6.87 (m, 1 H), 5.97 (s , 2 H), 3.49 (m, 2 H), 2.77 (m, 2 H).

Compound I-530 and reference compound I-531

The reference compound I-405 was dissolved by chiral separation of SRC with chiralcel AD-H 50 mm x 250 mm semipreparative column, using 15:85 ethanol+0.5% diethylamine:CO2. Collection of the two peaks and concentration in vacuo produced the reference compound I-530 (first elution peak by analytical HPLC, Chiralcel AD-H 4.6 mm x 250 mm, 15:85 ethanol+0.5% diethylamine :hexane) as a light orange solid. Collection of the second eluting peak and concentration in vacuo yielded reference compound I-531 (first eluting peak by analytical HPLC, Chiralcel AD-H 4.6 mm x 250 mm, 15:85 ethanol+diethylamine 0 .5%:hexane) as a light orange solid.

1H-NMR for Compound I-530 (500 MHz, DMSO-d6) 8 ppm 9.11 (d, 1 H), 8.32 (d, 1 H), 7.93 (t, 1 H), 7 .90 (s, 1 H), 7.78 (br s, 1 H), 7.69 (br s, 1 H), 7.51 (s, 1 H), 7.35-7.31 (m , 1 H), 7.22-7.19 (m, 2 H), 7.10 (t, 1 H), 7.00-6.97 (m, 1 H), 5.90 (s, 2 H), 4.02-3.94 (m, 2 H).

1H-NMR for Reference Compound I-531 (500 MHz, DMSO-d6) 8 ppm 9.11 (d, 1 H), 8.33 (d, 1 H), 7.93 (t, 1 H) , 7.90 (s, 1 H), 7.78 (br s, 1 H), 7.69 (br s, 1 H), 7.52 (s, 1 H), 7.35-7.31 (m, 1 H), 7.22-7.19 (m, 2 H), 7.10 (t, 1 H), 7.00 6.97 (m, 1 H), 5.90 (s, 2H), 4.02-3.94 (m, 2H).

Reference compound I-630

The title compound was synthesized in 2 steps:

Stage 1: Synthesis of (2-(1-(2-fluorobenzyl)-5-(isoxazol-3-yl)-1H-

methyl pyrazol-3-yl)pyrimidin-5-yl)carbamate

To a solution of 2-(1-(2-fluorobenzyl)-5-(isoxazol-3-yl)-1H-pyrazol-3-yl)pyrimidin-5-amine (this compound was described in the patent application publication WO2012003405 A1) (1 equiv.) In anhydrous pyridine at 0 ° C, methyl chloroformate (1.2 equiv.) was added. After stirring for 10 min, the reaction mixture was warmed to room temperature and carefully monitored by LC/MS. Additional parts of methyl chloroformate (3.2 equiv.) were added at 0 °C. After stirring at room temperature for 20 h, the crude mixture was diluted with water. The resulting tan solids were collected by filtration and used in the next step without further purification.

Stage 2: Synthesis of the reference compound I-630

A suspension of (2-(1-(2-fluorobenzyl)-5-(isoxazol-3-yl)-1H-

Methyl -pyrazol-3-yl)pyrimidin-5-yl)carbamate (1 equiv.) in DMF at 0 °C was treated with sodium hydride (60% w/w in mineral oil, 1.1 equiv.) and was warmed to room temperature. After 30 min, the reaction mixture was cooled to 0 °C and iodomethane (1.1 equiv.) was added. After 25 min, the crude reaction mixture was poured into water and extracted with ethyl acetate. The combined organic phases were dried over sodium sulfate, filtered, and the solvent was removed in vacuo. The crude material was purified by silica gel chromatography with a gradient of 20% acetonitrile:methanol (7:1) in dichloromethane providing the desired compound, reference compound I-630 (12 mg, 21% yield in two steps). like a white solid.

1H NMR (500 MHz, CD3OD) 5 ppm 8.88 (s, 2 H), 8.77 (d, 1 H), 7.55 (s, 1 H), 7.28 (ap. q, 1 H ), 7.09 (ap. t, 1 H), 7.04 (ap. t, 1 H), 6.91 (d, 1 H), 6.88 (ap. t, 1 H), 5, 99 (s, 2 H), 3.79 (s, 3 H), 3.40 (s, 3 H).

Reference Example 2A: Measurement of biological activity by sGC-HEK-cGMP assay (SNP incubation assay)

Human embryonic kidney cells (HEK293), endogenously expressing soluble guanylate cyclase (sGC), were used to evaluate the activity of the test compounds. Compounds that stimulate the sGC receptor should cause an increase in the intracellular concentration of cGMP. HEK 293 cells were seeded in Dulbecco's modification of Eagle's medium supplemented with fetal bovine serum (10% final) and L-glutamine (2 mm final) in a volume of 200 μl at a density of 1x105 cells/well in a plate. of 96-well flat-bottom cells coated with poly-D-lysine and cultured overnight at 37°C. The medium was aspirated and the cells were washed with (1x) Hank's buffered saline (200 μl). The cells were then incubated for 15 minutes at 37°C with 200 μl of a 0.5 mM 3-isobutyl-1-methylxanthine (IBMX) solution. Test article and sodium nitroprusside solutions were then added (x pM concentration for the test article solution and 10 pM concentration for the SNP solution; where x is one of the following concentrations: 30 pM, 10 pM , 3 pM, 1 pM, 0.3 pM, 0.1 pM, 0.03 pM, 0.01 pM, 0.003 pM, 0.001 pM, 0.0003 pM or 0.0001 pM) to the test mixture (2 pl each) and the resulting mixture was incubated at 37 °C for 10 minutes. After 10 minute incubation, the assay mixture was aspirated and 0.1 M HCl (200 μL) was added to the cells. The plate was incubated at 4 °C for 30 minutes in the 0.1 M HCl to stop the reaction and lyse the cells. The plates were then centrifuged at 1,200 g for 5 minutes at room temperature. The supernatants were collected and transferred to a new flat-bottom 96-well plate for HPLC-MS analysis. Vehicle controls were carried out with DMSO solutions (1%). A known sGC stimulator, BAY 41-2272, was used as a positive control. The samples were diluted with an equal volume of 1 M ammonium acetate (pH 7) to neutralize the samples for better chromatography. A 2x cGMP standard solution was prepared in 0.1 M HCl and then diluted with an equal volume of 1 M ammonium acetate, with the following final concentrations in nM: 1,024, 512, 256, 128, 64, 32, 16, 8, 4, 2, 1. The cGMP concentrations in the test plates were determined from each sample using the LC/MS conditions shown in Table 2 below and the calculated cGMP standard curve. EC50 values were calculated from concentration-response curves generated with GraphPad Prism software. Data were normalized to high control by using the following equation: 100 * (Sample - Low Control)/(High Control -Low Control), where Low Control is the average of 6 samples treated with 1% DMSO, and the high control is the average of 8-12 samples treated with 10uM BAY 41-2272. Data were fitted using nonlinear regression, sigmoidal dose response, 3 parameter settings. Samples were typically processed at n=1, but for samples that were processed at n=2 (or more) the results given here correspond to the arithmetic mean of the various results obtained for each given compound. If a curve did not plateau, then the 100% limitation was imposed.

Compounds that did not induce a minimum response of 30% were recorded as ND and EC50 values were not determined. The biological activities of some of the reference compounds determined with the sGC-HEK assay with SNP incubation are summarized in Table 3.

Table 2 (HPLC LC/MS experimental conditions)

Table 3. Whole cell activity in the HEK assay with LC/MS detection.

Example 2B: Measurement of biological activity by sGCs-HEK-cGMP assay (using HTRF detection) (SNP incubation assay)

Human embryonic kidney cells (HEK293), endogenously expressing soluble guanylate cyclase (sGC), were used to evaluate the activity of the test compounds. Compounds that stimulate the sGC enzyme should cause an increase in the intracellular concentration of cGMP. HEK 293 cells were seeded in Dulbecco's modification of Eagle's medium supplemented with fetal bovine serum (10% final) and L-glutamine (2 mm final) in a volume of 200 μl at a density of 1x105 cells/well in a plate. of 96-well flat-bottom cells coated with poly-D-lysine and cultured overnight at 37°C. The medium was aspirated and the cells were washed with (1x) Hank's buffered saline (200 μl). The cells were then incubated for 15 minutes at 37°C with 200 μl of a 0.5 mM 3-isobutyl-1-methylxanthine (IBMX) solution. The test article and sodium nitroprusside solutions were then added to the test mixture (2 μl each) and the resulting mixture was incubated at 37 °C for 10 minutes. After 10 minute incubation, the assay mixture was aspirated and 0.1 M HCl (200 μL) was added to the cells. The plate was incubated at 4 °C for 30 minutes in the 0.1 M HCl to stop the reaction and lyse the cells. The plates were then centrifuged at 1,200 g for 5 minutes at room temperature. GMP levels were determined using a cGMP HTRF assay (Cisbio Product #62GM2PEC). For each sample, 5ul of HEK assay supernatant was diluted 1:5 in HTRF kit assay diluent and transferred to a reservoir of the assay plate, and the HTR<f>assay was performed according to the kit manufacturer's instructions. HTRF. Sample calculations were performed using high and low controls, where the high control was supernatant from a HEK assay performed in the presence of 10 uM Bay 41-2272 SNP and the low control was the supernatant from the HEK assay performed in the presence of vehicle. A cGMP standard solution was prepared in 0.1 M HCl and diluted to perform a cGMP standard curve using the HTRF assay. Using mean ratio data from the HTRF assay, sample data were normalized according to the equation: 100 * (Sample - Low Control)/(High Control - Low Control). Data are fit to 3 log agonist-agonist dose response parameters (Top (%EMax), Bottom, log EC50) using Graphpad (Prism software). The data in Table 4 were obtained using this modified test procedure. (x pM concentration for the test article solution and 10 pM concentration for the SNP solution; where x is one of the following concentrations: 30 pM, 10 pM, 3 pM, 1 pM, 0.3 pM, 0.1 pM, 0.03 pM, 0.01 pM, 0.003 pM, 0.001 pM, 0.0003 pM and 0.01 nM.

Table 4. Whole cell activity in the HEK assay with HTRF detection.

Compound I-405 and Reference Compounds I-306 to I-404 and I-406 to I-455 were analyzed in this assay and most of them showed EC50 values less than 5.0 pM, with Emax values of at minus 80%.

Example 2 C: M e d i c i o n o f b i o l o g i c a c t i v i t i t y m e d i a n t e s s i o n G C - H E K - c G M P, new pr o t o c o l c o n d e t e c t i o n L C / M S

Human embryonic kidney cells (HEK293), endogenously expressing soluble guanylate cyclase (sGC), were used to evaluate the activity of the test compounds. Compounds that stimulate the sGC enzyme should cause an increase in the intracellular concentration of cGMP. HEK 293 cells were seeded in Dulbecco's modification of Eagle's medium supplemented with fetal bovine serum (10% final) and penicillin (100 U/ml)/streptomycin (100 p g/ml) in a volume of 50 μl at a density of 1. 5x104 cells/well in a 384-well flat-bottom plate coated with poly-D-lysine. Cells were incubated overnight at 37°C in a humidified chamber with 5% CO2. The medium was aspirated and the cells were washed with (1x) Hank's buffered saline (50 μl). The cells were then incubated for 15 minutes at 37°C with 50 μl of a 0.5 mM 3-isobutyl-1-methylxanthine (IBMX) solution. Next, test article and diethylenetriamine-NONOate (DETA-NONOate) solutions were added (concentration x pM for the test article solution and concentration 10 pM for the DETA-NONOate solution; where x is one of the following concentrations);

were added to the test mixture and the resulting mixture was incubated at 37 °C for 20 minutes. After 20 minutes of incubation, the assay mixture was aspirated and 10% acetic acid containing 150 ng/ml 3 cGMP (internal standard for LCMS) (50 μl) was added to the cells. The plate was incubated at 4 °C for 30 minutes in the acetic acid solution to stop the reaction and lyse the cells. The plates were then centrifuged at 1,000 g for 3 minutes at 4°C and the supernatant was transferred to a clean reaction plate for LCMS analysis.

The cGMP concentrations of each sample were determined using the LCMS conditions shown below (Table 5) and the standard curve was calculated. The standard curve was prepared in 10% acetic acid with 150 ng/ml 3cGMP (isotopically labeled cGMP 3 units higher than wild type) with the following final cGMP concentrations in ng/ml: 1, 5, 10, 50 , 100, 250, 500, 1000, 2000.

Table 5: LC/MS conditions, Example 2C

Data were normalized to high control by using the following equation: 100 * (Sample - Low Control)/(High Control - Low Control), where Low Control is the average of 16 samples treated with 1% DMSO, and the high control is the average of 16 samples treated with p30 M of I-329. Data were fitted using a 4-parameter fit (log (agonist) vs response - variable slope) with GraphPad Prism v software. 5. n=2 for all compounds. The absolute EC50 value was interpolated from the curve fit and is defined as the concentration at which a given compound elicits 50% of the high control response. Compounds that do not produce a minimum response of 50% are indicated > as 30 pM. For duplicate test compounds or n greater than 2, the result indicated here is the geometric mean of the different results obtained. Table 6 summarizes the results obtained for the compounds of the invention (I-405 and I-530) and the reference compounds selected in this test.

Table 6. Total cell activity in HEK assay with LC/MS detection (updated assay conditions,

Example 2C).

Reference Example 3A: Measurement of Biological Activity Using the Thoracic Aortic Ring Assay

The thoracic aortic rings are dissected from anesthetized male Sprague-Dawley (isoflurane) rats weighing 275,299 g. The tissues are immediately transferred to the very cold Krebs-Henseleit solution, which has been aerated with 95% O2 and 5% CO2 for 30 minutes. After removal of connective tissue, aortic sections are cut into 4 rings (~2 mm each) and suspended on 2 L-shaped hooks, with a fixed hook at the bottom of the tissue bath (Schuler Organ Bath, Harvard Apparatus) and the other connected to a force transducer (F30 force transducer, Harvard Apparatus). Baths containing Krebs Henseleit solution (10 mL) are heated to 37 °C and aerated with 95% O2 and 5% CO2. The rings are brought to an initial tension of 0.3-0.5 g and gradually increased to a resting tension of 1.0 g over 60 minutes. The rings are rinsed with Krebs Henseleit solution (heated to 37°C and aerated with 95% O2 and 5% CO2) at 15 minute intervals until a stable baseline is obtained. The rings are considered stable after maintaining a resting tension of 1.0 g (for approximately 10 minutes) without adjustment. The rings are then contracted with 100 ng/ml phenylephrine by adding 100 ul of a 10 pg/ml phenylephrine stock solution. Tissues that achieve stable contraction are then treated in a dose-dependent, cumulative manner with test compounds prepared in dimethyl sulfoxide (DMSO). In some cases, tissues are rinsed three times over a period of 5 minutes with Krebs-Heinseleit solution (warmed to 37°C and aerated with 95% O2 and 5% CO2), allowed to stabilize at baseline, and They are then used for the characterization of other test articles or effects of DMSO. All data are taken with HSE-ACAD software provided by Harvard Apparatus. Percent relaxation effects are calculated in Microsoft Excel using the recorded tension value of 100 ng/ml phenylephrine treatment as 0% inhibition and 100 pM 3-isobutyl-1-methylxanthine treatment as 100% inhibition. EC50 values are calculated from concentration response curves generated with GraphPad Prism software.

Reference Example 3B: Measurement of Biological Activity Using the Thoracic Aortic Ring Assay, Alternative Method

As an alternative thoracic aortic ring assay, the procedure of Reference Example 3 is used, except that the percent relaxation effects are calculated in Microsoft Excel using the recorded 100 ng/ml phenylephrine treatment tension value as 0% inhibition. and, after washing the fabric with buffer, the original resting tension of the fabric is used as 100% inhibition.

Example 4: Change in blood pressure in Sprague-Dawley rats

Male rats (250-350 g body weight, supplied by Harlan Laboratory) were anesthetized. s c o n k e t a m i n a / x i l a z i n a and a c a t e t e r g a d o f sal i n o h e p a r i n i z a n d i m p l a n t e d i n t h e right f e m o r a l a r t e r . The catheter was externally placed between the scapula, plugged, and the animal was allowed to recover for at least 7 days after u e of the surgery before performing any compositing tests. Before the test, the animals were kept on a normal diet, with free access to drinking water, in a cycle of light. 12 h o r a s .

On the experimental day, under anesthesia with inhaled isoflurane, the catheter was uncapped and connected to a cable (Instech Labs) and pressure transducer (Harvard Apparatus). Subsequently, their blood pressure and heart rate were taken and analyzed with a data capture system used for this (PowerLab, ADInstruments). Data sampling rates were set to 1 cycle per second. Once connected, each rat was allowed to recover from anesthesia and baseline blood pressure and heart rate levels were established in these conscious and free-to-move animals. Once the baseline value was established, either vehicle (0.5% methylcellulose or PEG400100%) or test article was administered orally (PO, 10 mg/kg) and the effects on blood pressure and rhythm were monitored. cardiac for 24 hours.

Data are reported as hourly averages and changes in blood pressure are calculated by subtracting individual baseline values each hour.

Reference Example 5: Animal Model Descriptions:

Lamb model of hemodynamics using inhaled sGC stimulator

(“Inhaled Agonists of Soluble Guanylate Cyclase Induce Selective Pulmonary Vasodilation”, Oleg V. et al., American J of Respons and Critical Care Medicine, vol. 176, 2007, p. 1138)

It is possible to test whether inhalation of novel formulations of dry powder microparticles containing sGC stimulators would produce selective pulmonary vasodilation in lambs with acute pulmonary hypertension following a published procedure. It is also possible to evaluate the combined administration of sGC stimulator microparticles and inhaled nitric oxide (iNO) in this system. Finally, it is possible to examine whether inhalation of microparticles from a G C stimulator would produce pulmonary vascular dilation when affected by t a a i N O (s i n t a s a i n d u c i b l e ) .

Protocol: In spontaneously breathing, awake lambs with implanted vascular catheters and a tracheostomy tube, U-46619 is insufflated intravenously to increase pulmonary arterial pressure to 35 mm Hg. Inhalation of microparticles composed of BAY 41-2272, BAY 41-8543, or Ba Y 58-2667 and excipients (dipalmitoylphosphatidylcholine, albumin, lactose) produced dose-dependent pulmonary vasodilation and an increase in transpulmonary cGMP release without a significant effect in mean arterial pressure. Inhalation of microparticles containing BAY 41-8543 or BAY 58-2667 increased systemic arterial oxygenation. The magnitude and duration of iNO-induced pulmonary vasodilation increased after inhalation of BAY 41-8543 microparticles. Intravenous administration of 1H-[1,2,4]oxadiazolo[4,3-a]quinoxalin-1-one (ODQ), which oxidizes the prosthetic heme group of sGC, markedly reduced the pulmonary vasodilatory effect of iNO. In contrast, pulmonary vasodilation and transpulmonary cGMP release induced by inhalation of BAY 58-2667 microparticles were greatly increased after ODQ treatment. Therefore, inhalation of microparticles containing sGC agonists may provide an effective novel treatment for patients with pulmonary hypertension, especially when iNO responsiveness is affected by sGC oxidation. Note: BAY 41-2272, Ba Y 41-8543 are sGC stimulators, while BAY 58-2667 is an sGC activator.

In vitro (ex vivo) model of guinea pig tracheal smooth muscle stimulated with an electric field for the evaluation of bronchodilation.

It is possible to evaluate the bronchodilator effects of sGC stimulators using the system described below. This system allows us to determine the potency, efficacy and duration of action of various sGC stimulators, as well as evaluate potential adverse effects such as changes in blood pressure or heart rate.

Animals: Guinea pigs, Dunkin Hartley, male, unrestricted feeding and certified free of specific microorganisms at the time of receipt of 525-609 g on the day of the experiment. Harlan UK Ltd. guinea pigs are housed in a group of 4 in solid bottom cages with Gold Flake bedding in a controlled environment (airflow, temperature and humidity). Food (FD1, Special Diet Services) and water are provided ad libitum.

Contraction of guinea pig tracheal smooth muscle in response to EFS. Evaluation of the potency and effectiveness of the compound:

On each day of experiments, one guinea pig is killed by exposure to an increasing concentration of CO2 and removal from the trachea. The trachea is cleared of foreign tissue and opened by cutting it longitudinally in a line in front of the muscle, opened and cut into strips a width of 2-3 rings of cartilage. A loop of cotton is attached to one end of each tracheal strip and a length of cotton to the other end. The tracheal strips are then suspended between two platinum electrodes, using tissue supports, in a Myobath system (World Precision Instruments Stevenage, UK). The loop is attached over the hook at the bottom of the tissue holder and the other end is attached to the arm of a FORT10 force transducer (World Precision Instruments Stevenage, UK) ensuring that the tissue is positioned between the two platinum electrodes. . The entire unit is then lowered into a 10 ml tissue bath containing Kreb's modified Henseleit buffer at 37°C, with carbogen sparging. A tension of 1 g is applied to each piece of fabric and the fabric is washed, followed by a 1 hour stabilization period. Once the tissues have been allowed to stabilize, the electric field stimulation apparatus is configured to deliver 80 Hz frequency stimulation of 0.1 ms pulse width, with a switched, unipolar pulse every 2 minutes using a DS8000 8-channel digital simulator (World Precision Instruments Stevenage, UK). A voltage response curve is carried out on each strip at 2, 4, 6, 7, 8, 10, 12 V and then a submaximal voltage is selected to apply to each tissue for the remainder of the experiment. Contraction of guinea pig tracheal smooth muscle (GPTSM) is induced using submaximal Electrical Field Stimulation (EFS) (it is also possible to induce contraction using a spasmogenic substance, such as methacholine or histamine as described in Coleman et al.*). Compounds are dissolved in 100% DMSO at 3x10-2 M and aliquots are stored at -200°C. A separate aliquot is used for each experiment. Tissues are washed with Krebs buffer and stimulated using the previously determined submaximal voltage for 1 hour to establish a stable baseline contraction prior to evaluation of compound activity.

A cumulative dose response curve (DRC) is then performed for each test substance and changes in smooth muscle contraction are measured. The effect of each test substance in each experiment is expressed as percentage inhibition of base contraction, normalized to the relevant vehicle controls. The experiment is performed three times using tissue from three different animals. The data from the three experiments are combined, the DRC is plotted, and the potency of the test substance and the efficacy are determined. The potency of ipratropium bromide is evaluated along with the test compounds and the IC50 value determined is 0.86 nM (95% Cl, 0.78-0.94), based on the data previously obtained in the system.

“Novel and Versatile Superfusion System. Its use in the Evaluation of Some Spasmogenic and Spasmolytic Agents Using Guinea pig isolated Tracheal Smooth Muscle.” , R. A. Coleman et al., J. Pharmacol. Methods, 21, 71-86, 1989.

Mouse model for diseases in which altered CFTR function is causally involved.

These diseases include c h i s t i c fi b r o s i n a l e s , liv er disor ders , d i a b e t e s rel a t i o n a d a c o n c h i s t i c f i b r o s i e (C F R O ) e S j o e g r e n .

Using transgenic mice that express or do not express the F508CFTR delta channel it is possible to measure differences in nasal potential difference and salivation in the presence of a test sGC stimulator using the literature protocol described below (see WO2011095534).

Salivary secretion assay in delta(.6.)50S-CFTR mice

15 male and female homozygous, 6.50S-CFTR heterozygotes (backcrossed on the FVB genetic background over 12 generations, originally obtained from Erasmus University Rotterdam; those used in this trial were 10-14 weeks old and weighed 1S-36 g , of both sexes Vardenafil solutions were prepared at concentrations of 0.07, 0.14 and 0.42 mg/kg BW in sterile saline, while the sGC stimulator BAY 41-2272 was dissolved in 0.01, 0.03, 0.1 and 0.3 mg/kg BW in a solvent containing ddH2O, 40% PEG 400 (polyethylene glycol 400) and 10% ethanol The appropriate substances or vehicles were administered to mice by intraperitoneal injection (5. ml/kg body mass) 60 min before the salivary secretion test. After 60 min, the mice were anesthetized with a combination of ketamine and diazepam. The solution was prepared to contain 1 ml of 5 mg/diazepam. ml and 1 ml of 100 mg/ml ketamine in 8 ml of sterile saline. Anesthesia was induced by intraperitoneal injection of the solution (10 ml/kg body mass). After anesthesia, mice were pretreated with a subcutaneous injection of 1 mM atropine (50 1-11) in the left cheek to avoid cross-stimulation of cholinergic receptors. Small strips of Whatman 5 filter paper were placed inside the previously injected cheek for 4 min to absorb saliva secreted after atropine injection. This first piece of filter paper was removed and replaced with a second, pre-weighed filter paper. After that, 50 1-11 of a solution containing 100 I-IM isoprenaline and 1 mM atropine was injected into the left cheek at the same site to induce salivary secretion by adrenergic mechanisms. The time of isoprenaline injection 10 was taken as time zero, and the filter paper strips were replaced every 10 minutes for a total collection period of 30 minutes. Each piece of filter paper was immediately placed and sealed in a pre-weighed vial. After all samples were collected, each vial was measured again and the weights of all samples were recorded. The difference in the total weight of the vial plus paper measured before and after collecting saliva was taken as the net weight of saliva secreted during the collection period. Total amounts of salivary secretion were calculated as the weight of saliva divided by the number of minutes required for each collection and then normalized according to the mass of the mouse in grams. Results are expressed as the mean percentage increase in n mice compared to placebo treatment. Statistical analysis was performed using a one-way ANOVA followed by a post-hoc Bonferroni analysis; *****//* means statistically significant with p values <0.05/<0.01/0.001 and n.s. means not significant.

These animal studies were carried out with various sGC stimulators, sGC activators, and PDE5 inhibitors. The results suggest that the compounds of the invention are useful for the treatment of cystic fibrosis, pancreatic disorders, gastrointestinal disorders, liver disorders, Cystic Fibrosis Related Diabetes (CFRO), dry eye, dry mouth and Sjoegren's syndrome.

Neuromuscular disorders

Mislocalization of nitric oxide synthase (nNOS) from the sarcolemmal membrane to the sarcoplasm has previously been shown to be observed in a wide range of non-dystrophic neurological conditions associated with impaired motility status and catabolic stress. One tool for the evaluation of muscle biopsies from patients with various inherited and acquired forms of neuromuscular disorders is the evaluation of sarcolemmal localization of nNOS. The level of nNOS in the sarcolemma was found to correlate with mobility and functional status.

An analogous assessment can be used to determine nNOS localization in animal models of non-dystrophic myopathy following the literature protocols described below (“Loss of sarcolemmal nNOS is common in acquired and inherited neuromuscular disorders”; E.L. Finanger Heddericky al., Neurology,2011, 76(11), 960-967).

Mislocalization of nNOS in mouse models of acquired muscle atrophy

Two mouse models have been described that demonstrate muscle atrophy without compromising mobility: short-term unfed and high-dose corticosteroid therapy. Mice treated with steroids or not fed for 48 hours showed significant decreases in overall body mass and normalized wet skeletal muscle mass. Morphometric analysis of skeletal muscle specimens from both models showed muscle atrophy, defined by a significant decrease in minimum Feret fiber diameter compared to age-adjusted controls (n = 5 for each group). However, immunofluorescence staining for dystrophin, α-sarcoglycan, and α-1-syntrophin showed dystrophin localization suggestive of intact DGC complex. However, both steroid-treated and unfed mice had no or greatly reduced sarcolemmal nNOS staining. Real-time PCR for NOS family proteins (nNOS, eNOS, iNOS) did not reveal significant differences in the expression levels of any of the 3 transcripts in the mice studied with steroids (n = 8 for each group). . In addition, the Western blot analysis for N O S, N O S, and N O S did not show differences in the protein levels.

These murine animal models could be used to evaluate the effects of sGC stimulators (e.g., an sGC stimulator of the invention) on symptoms of muscle atrophy and related disease states.

Unfed mice showed a 100% decrease in nNOS and iNOS transcript expression compared to wild-type mice (n = 9 for controls, n = 7 for unfed). However, the protein level of nNOS, iNOS, and eNOS revealed no differences between control and unfed mice (n = 4 for each group). These data demonstrate that abnormal localization of nNOS occurs in mice with severe muscle atrophy even if overall mobility is preserved, supporting the notion that, in addition to decreased mobility, other triggering factors, such as catabolic stress, may be associated with sarcolemmal loss of nNOS.

Skeletal muscle nNOS localization is maintained during hibernation (squirrel studies)

Skeletal muscle specimens from hibernating thirteen-banded ground squirrels have been used to evaluate the effect of immobility and catabolic stress on nNOS localization in the context of maintained muscle homeostasis and integrity. These animals are necessarily hibernating mammals that are protected from musculoskeletal atrophy during hibernation. Despite hibernating for 5 months with almost complete immobility and no caloric intake, sarcolemmal expression of nNOS is maintained. These data together with data from patients and mice indicate that biochemical control of nNOS localization is complex and, importantly, conservation of sarcolemmal nNOS may be significant in maintaining muscle homeostasis.

These results also suggest that targeting aberrant NO signaling (e.g., with sGC stimulators such as those described here) may be beneficial for a broad group of patients with neuromuscular disorders.

Mouse models of Muscular dystrophy (BMD and DMD)

Becker muscular dystrophy (BMD), characterized by progressive wasting of skeletal muscle, is caused by mutations of the muscle protein dystrophin. In a human study, Martin et al. (see “ Tadalafil Alleviates Muscle Ischemia in Patients with Becker Muscular Dystrophy” ; Elizabeth A. Martin et al., Sci. Transí. Med.

4, 162ra155 (2012); “Vascular-targeted therapies for Duchenne muscular dystrophy”; Ennen et al., Skeletal Muscle, 2013, 3:9) evaluated exercise-induced attenuation of reflex sympathetic vasoconstriction in the muscles of 10 BMD patients and healthy male controls age-adjusted to 7 years. This is a protective mechanism that optimizes skeletal muscle perfusion to meet the metabolic demands of exercise. Vasoconstriction was induced by simulated orthostatic stress and was measured when the forearm muscles were at rest or lightly exercised in a rhythmic grip fashion. First, the researchers showed that exercise-induced attenuation of reflex vasoconstriction was deficient in 9 of 10 BMD patients in whom common dystrophin mutations disrupt neuronal NO synthase (nNOS) signaling to the muscle sarcolemma. Next, in a double-blind, randomized, placebo-controlled crossover trial, the authors showed that normal blood flow regulation was restored in eight of nine patients with a single oral dose of 20 mg of tadalafil, a specific PDE5 inhibitor.

It is possible to evaluate the effects of drugs that act on the NO pathway using a model of the related disease Duchenne muscular dystrophy (DMD) for dystrophin-deficient mice. This model has also shown that phosphodiesterase 5 (PDE5) inhibitors alleviate some features of the dystrophic phenotype, including vasospasm of skeletal muscle microvessels that can lead to muscle injury and fatigue.

With exercise of healthy skeletal muscle, sarcolemmal nNOS-derived NO attenuates local adrenergic vasoconstriction, thereby optimizing perfusion to meet the metabolic demands of active muscle. This protective mechanism (termed functional sympatholysis) is lost in mdx mice (a model of BMD and DMD), mice without nNOS, and boys with DMD, causing functional muscle ischemia. Repeated attacks of functional ischemia could accelerate use-dependent damage to muscle fibers already weakened by dystrophin deficiency.

In the mdx mouse, many features of the dystrophic phenotype can be ameliorated by multiple strategies that drive NO signaling, including transgenic expression of dystrophin minigenes that restore sarcolemmal nNOS (and thus restore functional sympatholysis), administration of the NOS substrate L -arginine (24, 25), treatment without NO donor drugs, and inhibition of phosphodiesterase 5A (PDE5A) with the drug tadalafil or sildenafil. These PDE5A inhibitors, which prolong the half-life of guanosine-3',5'-monophosphate (cGMP)—the downstream target of NO in vascular smooth muscle—were shown to alleviate muscle ischemia, as well as injury and death, in the mouse. fatigue after a short exercise session. Also, these drugs were shown to improve cardiac dynamics in mice mdxy rescue dystrophic skeletal muscle and prolong survival in dystrophin-deficient zebrafish.

These results support an essential role for sarcolemmal nNOS in modulating sympathetic vasoconstriction in human skeletal muscles and suggest that targeting the aberrant NO pathway (e.g. using an sGC stimulator of the invention) may be a useful therapeutic approach. to treat BMD and DMD in humans.

Sickle cell disease

Sickle cell disease (SCD), or sickle cell anemia (SCA) or sickle cell disease, is an inherited blood disorder, characterized by red blood cells that become “sickle” (sickle) shaped, rigid and abnormal. Sickle cell disorder decreases the flexibility of cells and results in the risk of various complications. Sickle cell disorder occurs due to a mutation in the hemoglobin gene. Individuals with one copy of the silent gene show both normal and abnormal hemoglobin. This is an example of co-dominance. In 1994 in the US, the average life expectancy of people with this condition was estimated to be 42 years for men and 48 years for women, but today, thanks to With better disease management, patients can live 70 years or more.

Sickle cell anemia is a form of sickle cell disease in which there is homozygosity for the mutation that causes HbS. Sickle cell anemia is also called “HbSS,” “SS disease,” “hemoglobin S,” or permutations of those names. In heterozygous people, that is, those who have only one sickle cell gene and one normal adult hemoglobin gene, the condition is called “HbAS” or “sickle cell trait.” Other less common forms of sickle cell disease are compound heterozygous states in which the person has only one copy of the mutation that causes HbS and one copy of another abnormal hemoglobin allele. They include hemoglobin C sickle cell disease (HbSC), sickle beta plus thalassemia (HbS/p+), and sickle beta zero thalassemia (HBS/p°).

Although sickled red blood cells (RBCs) and rheological abnormalities are highly characteristic of the pathophysiology of sickle cell disease, vascular dysfunction resulting from complex interactions between sickled red blood cells (sRBCs), endothelial cells, platelets, and leukocytes play an equally important role. In sickle cell disease, endothelial activation is associated with sickle cell-mediated hypoxiareperfusion events (see, for example, “Advances in understanding of the pathogenesis of cerebrovascular vasculopathy in sicke cell anemia,” P. Connes et al., Br. . J. Haematol.2013, l6 l, 484-98). Sickle-shaped deformation of red blood cells and adhesion to the endothelium initiate vasoclusion, affecting blood flow. The subsequent increase in inflammatory mediators and endothelial activation trigger a cascade of events that produce vascular damage. Pathophysiological responses to intermittent hypoxia-reperfusion from these vaso-occlusive events are manifested by increased cytokine production, leukocyte regulation, and activation of procoagulant and adhesion molecules, with simultaneous inhibition of cytoprotective mediators.

Leukocytosis correlates with almost all manifestations of sickle cell disease, underscoring the important role of inflammation in the pathophysiology of sickle cell disease. Even at baseline, patients with sickle cell disease have elevations in proinflammatory cytokines, including C-reactive protein (CRP), tumor necrosis factor (TNF), interleukin-1 (IL-1), and interleukin-8. (IL-8). In vitro studies have shown that sRBCs promote endothelial positive regulation of TNF-a and IL-1-p (8-1°). Microarray studies of activated monocytes have demonstrated differential expression of genes involved in inflammation, heme metabolism, cell cycle regulation, antioxidant responses, and angiogenesis. More recently, differential expression of nuclear factor kappa light chain enhancer of activated B cells (NFKB/p65), Kruppel-like factor 2 (KLF2), and other transcription factors has been shown to regulate inflammatory pathways. in children with sickle cell disease with increased risk of stroke.

In transgenic mouse models (see “Novel Therapies Targeting the Endothelium in sickle cell disaease”, C.C Hoppe, Hemoglobin, 35(5-6):530-546 (2011) and the references cited therein), it has been shown that Oxidative stress that produces sickle cell deformation alters microvascular regulatory mechanisms leading to endothelial activation and exaggerated inflammatory and proadhesive responses. Oxidative stress occurs through the formation of reactive oxygen species (ROS). NO depletion occurs through hemoglobin (Hb)-mediated removal, ROS consumption, and arginase-mediated substrate depletion. In sickle cell disease the scavenging systems that normally remove freely circulating Hb are saturated. Free Hb depletes NO, leading to endothelial dysfunction. Therefore, the normal balance of vasoconstriction and vasodilation tends toward vasoconstriction, endothelial activation, oxidative stress, and proliferative vasculopathy.

Therapies aimed at restoring NO homeostasis have shown promise in preliminary studies in patients with sickle cell disease. Previous in vitro studies and studies in other patient populations showed NO-mediated downregulation of endothelial adhesion molecule expression. Following these observations, the use of inhaled NO was studied in children with sickle cell disease who had VOE and found trends associated with lower pain scores, lower analgesic requirements, and shorter hospital stay.

These findings were replicated in a recent randomized placebo-controlled trial evaluating inhaled NO for the treatment of acute<v>O<e>in adult patients with sickle cell disease, showing that inhaled NO significantly reduced pain scores and was associated to a trend toward decreased use of parenteral morphine compared with placebos. Results from a full phase II trial of patients with sickle cell disease treated with inhaled NO for acute VOE are not yet available (clinicaltrials.gov NCT00023296). Another phase II trial of inhaled NO for the treatment of VOE in children with sickle cell disease is expected to be completed (clinicaltrials.gov NCT00094887). The possible therapeutic role of inhaled NO for ACS in sickle cell disease is currently being evaluated in both children and adults in two different French phase II/III trials comparing the use of inhaled NO with placebo or care standard in children with ACS (clinicaltrials.gov NCT01089439 and NCT00748423).

Dietary supplementation of the NO synthase substrate, L-arginine, has been studied extensively in sickle cell disease as a means of increasing NO bioavailability. In mice with sickle cell disease, L-arginine at high doses has been shown to decrease Gardos canal activity, hemolysis and dense cell formation, as well as functional improvements in vascular reactivity.

Sildenafil, an agent aimed at amplifying the effect of endogenous NO by inhibiting PDE5, a downstream mediator of NO, is widely used in the general population to treat primary PHT. Preliminary studies in sickle cell disease patients with severe PHT described improvements in PAP and exercise capacity after treatment with sildenafil. A multicenter trial (Treatment of Pulmonary Hypertension and Sickle Cell Disease with Sildenafil Therapy, Walk-PHaSST) studying the safety and efficacy of sildenafil in patients with sickle cell disease with Doppler-defined PHT was terminated prematurely due to increased frequency of serious side effects, including increased VOE velocities, headache, and visual disturbance in the treatment group.

Nitrite and niacin have also been investigated for their properties as direct NO donors. In a phase I/II clinical trial, sodium nitrite infusions in patients with sickle cell disease improved forearm blood flow, consistent with an NO donor mechanism of action. A larger phase I/II trial is investigating whether administering nitrite infusions as adjuvant therapy during acute VOE will improve blood flow and tissue oxygenation (clinicaltrials.gov NCT01033227). The effect of niacin on endothelium-dependent vasodilation is also being evaluated in a phase II randomized controlled trial (clinicaltrials.gov NCT 00508989).

The above results suggest that targeting the aberrant NO pathway in sickle cell disease (e.g., using an sGC stimulator of the invention) may be a useful therapy for the treatment of the disease. Murine models of sickle cell anemia that could be used to evaluate the effect of sGC stimulators (e.g., an sGC stimulator of the invention) in this disease state are described in Blood, 2001, 98(5), 1577- 84;J. Clin. Invest.2004, 114(8), 1136-45; yBr. J. Haematol.,2004, 124(3), 391-402.

Bladder dysfunction

The sGC activator BAY 60-2770 has been shown to improve overactive bladder syndrome in obese mice (see “The Soluble Guanylyl Cyclase Activator BAY 60-2770 ameliorates overactive bladder in obese mice”, Luiz O Leiria et al., The Journal of Urology,2013, doi:10.1016/j.juro.2013.09.020.). The animal model described in this publication can be used analogously to evaluate the effect of an sGC stimulator (for example, an sGC stimulator of the invention) in overactive bladder syndrome.

The same group of researchers has also described a rat model of bladder dysfunction (NO-deficient rats, F Z Monica et al., Neurology and Urodynamics, 30, 456-60, 2011) and have shown the protective effects of BAY -2272 (an sGC activator) in this model. The animal model described in this publication can be used analogously to evaluate the effect of an sGC stimulator (e.g., an sGC stimulator of the invention) on bladder dysfunction related to detrusor smooth muscle overactivity.

Claims (14)

CLAIMS 1. A compound represented by the following structural formula: or a pharmaceutically acceptable salt thereof. 2. A compound represented by the following structural formula: or a pharmaceutically acceptable salt thereof. 3. A pharmaceutical composition comprising a compound of claim 1 or 2, or a pharmaceutically acceptable salt thereof, and one or more excipients. 4. A compound of claim 1 or claim 2, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition of claim 3 for use in the treatment of a disease, health condition or disorder selected from the group consisting of: (1) peripheral, pulmonary, hepatic, kidney, cardiac or cerebrovascular/endothelial disorders/conditions or diseases otherwise related to circulation selected from: • disorders related to high blood pressure and reduced coronary blood flow; increased acute and chronic coronary blood pressure; arterial hypertension; vascular disorder due to cardiac and renal complications, heart disease, stroke, cerebral ischemia or renal failure; resistant hypertension; diabetic hypertension; congestive heart failure; diastolic or systolic dysfunction; coronary insufficiency; arrhythmias; reduction of ventricular preload; cardiac hypertrophy; heart/cardiorenal failure syndrome; portal hypertension; endothelial dysfunction or injury; • thromboembolic disorders and ischemia; myocardial infarction; ictus; transient ischemic attacks (TIA); obstructive thromboanginitis; stable or unstable angina pectoris; coronary spasms; variant angina; Prinzmetal's angina; restenosis resulting from thrombolysis therapies; thrombogenic disorders; • Alzheimer disease; Parkinson's disease; dementia; vascular cognitive impairment; cerebral vasospasms; head trauma; • Peripheral arterial disease; occlusive peripheral arterial disease; peripheral vascular disease; hypertonias; Raynaud's syndrome or phenomenon; critical limb ischemia; vasculitis; peripheral embolism; intermittent claudication; vaso-occlusive crises; Duchenne muscular dystrophy; Becker muscular dystrophy; microcirculation abnormalities; problems with exudation or vascular permeability; • shock; septicemia; cardiogenic shock; control of leukocyte activation; inhibition or modulation of platelet aggregation; • lung/respiratory conditions; pulmonary hypertension; pulmonary arterial hypertension and associated pulmonary vascular remodeling; localized thrombosis and right cardiac hypertrophy; pulmonary hypertonia; primary pulmonary hypertension; secondary pulmonary hypertension; familial pulmonary hypertension; sporadic pulmonary hypertension, precapillary pulmonary hypertension; idiopathic pulmonary hypertension; thrombotic pulmonary arteriopathy; plexogenic pulmonary arteriopathy; cystic fibrosis; bronchoconstriction or pulmonary bronchoconstriction; acute respiratory distress syndrome; pulmonary fibrosis; lung transplant; • pulmonary hypertension associated with or related to left ventricular dysfunction, hypoxemia, hypertension of WHO groups I, II, III, IV and V, mitral valve disease, constrictive pericarditis, aortic stenosis, cardiomyopathy, mediastinal fibrosis, pulmonary fibrosis , anomalous pulmonary venous drainage, pulmonary veno-occlusive disease, pulmonary vasculitis, collagen vascular disease, congenital heart disease, pulmonary venous hypertension, interstitial lung disease, sleep disordered breathing, sleep apnea, alveolar hypoventilation disorders, chronic exposure to high altitude, disease pulmonary in neonates, alveolar capillary dysplasia, sickle cell disease; coagulation disorders; chronic thromboembolism, pulmonary embolism due to tumor, parasites or foreign material, connective tissue disease, lupus, schistosomiasis, sarcoidosis, chronic obstructive pulmonary disease, asthma, emphysema, chronic bronchitis, pulmonary capillary hemangiomatosis; histiocytosis • atherosclerotic diseases or conditions; atherosclerosis; atherosclerosis associated with endothelial injury, platelet and monocyte adhesion and aggregation, smooth muscle proliferation and migration; restenosis; restenosis developed after thrombolysis therapies, percutaneous transluminal angioplasties (PTA), percutaneous transluminal coronary angioplasties (PTCA), and bypasses; inflammation; • cardiovascular disease associated with metabolic syndrome, obesity, dyslipidemia, diabetes, high blood pressure; lipid-related disorders, such as dyslipidemia, hypercholesterolemia, hypertriglyceridemia, sitosterolemia, fatty liver disease, and hepatitis; preeclampsia; progression of polycystic kidney disease; subcutaneous fat accumulation; • hepatic cirrhosis; liver cirrhosis associated with chronic liver disease; liver fibrosis, activation of hepatic stellate cells, hepatic fibrous collagen; total collagen accumulation; liver disease of necroinflammatory and/or immunological origin; • disorders of the urogenital system; kidney fibrosis; kidney failure resulting from chronic kidney disease or failure; renal failure due to tissue accumulation/deposition and injury, progressive sclerosis and glomerulonephritis; prostatic hypertrophy; • systemic sclerosis; • cardiac interstitial fibrosis; cardiac remodeling and fibrosis; cardiac hypertrophy; (2) ischemia or reperfusion injury; ischemia/reperfusion associated with organ transplant, lung transplant, lung transplant, or heart transplant; preservation of blood substituents in trauma patients; (3) a selected sexual, gynecological, and urological disorder of erectile dysfunction; impotence; premature ejaculation; female sexual dysfunction; female sexual desire dysfunction; hypoactive sexual desire disorder; vaginal atrophy; dyspaneuria; atrophic vaginitis; benign prostatic hyperplasia (BPH) or benign prostatic hypertrophy or enlargement; bladder outlet obstruction; painful bladder syndrome (BPS); interstitial cystitis (IC); overactive bladder, neurogenic bladder and incontinence; diabetic nephropathy; (4) ocular diseases or disorders selected from glaucoma, retinopathy, diabetic retinopathy, blepharitis, dry eye syndrome, Sjogren's syndrome; (5) hearing diseases or disorders selected from hearing impairment; partial or total hearing loss; partial or total deafness; tinnitus; noise-induced hearing loss; (6) selected topical or skin disorders or conditions of dermal fibrosis, scleroderma, skin fibrosis; (7) wound healing; wound healing in diabetics; improvement of microvascular perfusion; improving microvascular perfusion after injury, to counteract the inflammatory response in perioperative care; anal fissures; diabetic ulcers; and (8) other diseases or conditions selected from cancer metastasis; osteoporosis, gastroparesis; functional dyspepsia; diabetic complications; diseases associated with endothelial dysfunction; and neurological disorders associated with decreased nitric oxide production. 5. The compound, pharmaceutically acceptable salt or pharmaceutical composition for use according to claim 4, wherein the disease, condition or health disorder is a peripheral, pulmonary, vascular/endothelial disorder or condition, liver, kidney, heart, brain, or a disease related in another way to the circulation. 6. The compound, pharmaceutically acceptable salt pharmaceutical composition for use according to claim 4, wherein the disease, condition or health disorder is selected from ischemia, reperfusion injury, ischemia / reperfusion associated with organ transplantation, transplantation of lung, lung transplant or heart transplant, and preserving blood substituents in trauma patients. 7. The compound, pharmaceutically acceptable salt or pharmaceutical composition for use according to claim 4, wherein the disease, condition or health disorder is a sexual, gynecological or urological disorder or condition selected from erectile dysfunction; impotence; premature ejaculation; female sexual dysfunction; female sexual desire dysfunction; hypoactive sexual arousal disorder; vaginal atrophy, dyspareunia, atrophic vaginitis; benign prostatic hyperplasia (BPH) or hypertrophy or enlargement; bladder outlet obstruction; painful bladder syndrome (BPS); interstitial cystitis (IC); overactive bladder, neurogenic bladder and incontinence; and diabetic nephropathy. 8. The compound, pharmaceutically acceptable salt or pharmaceutical composition for use according to claim 4, wherein the disease is an ocular disease or disorder selected from glaucoma, retinopathy, diabetic retinopathy, blepharitis, dry eye syndrome and Sjogren's syndrome . 9. The compound, pharmaceutically acceptable salt or pharmaceutical composition for use according to claim 4, wherein the disease is a hearing disease or disorder selected from hearing deficiency, partial or total hearing loss; partial or total deafness; tinnitus; and noise-induced hearing loss. 10. The compound, pharmaceutically acceptable salt or pharmaceutical composition for use according to claim 4, wherein the disease is a topical or cutaneous disorder or condition selected from dermal fibrosis, scleroderma and cutaneous fibrosis. 11. The compound, pharmaceutically acceptable salt or pharmaceutical composition for use according to claim 4, wherein the treatment involves wound healing; wound healing in diabetics; improvement of microvascular perfusion; improvement of microvascular perfusion problems after injury; treatment of anal fissures; and treatment of diabetic ulcers. 12. The compound, pharmaceutically acceptable salt or pharmaceutical composition for use according to claim 4, wherein the disease, or condition is selected from cancer metastasis; osteoporosis; gastroparesis; functional dyspepsia; diabetic complications; diseases associated with endothelial dysfunction; and neurological disorders associated with decreased nitric oxide production. 13. The compound, pharmaceutically acceptable salt or pharmaceutical composition for use according to any one of claims 4-12, wherein the compound is used in combination with one or more additional therapeutic agents. 14. The compound, pharmaceutically acceptable salt or pharmaceutical composition for use according to claim 13, wherein one or more additional therapeutic agents are selected from endothelium-derived releasing factor, NO donors, substances that improve cGMP concentrations, nitric oxide synthase substrates, compounds that enhance eNOS transcription, heme-independent sGC activators, heme-dependent sGC stimulators, cGMP degradation inhibitors, calcium channel blockers, endothelin receptor antagonists, prostacyclin derivatives, antihyperlipidemics, anticoagulants, antiplatelet drugs, ACE inhibitors, supplemental oxygen, beta blockers, antiarrhythmic agents, diuretics, exogenous vasodilators, bronchodilators, corticosteroids, dietary supplements, PGD2 receptor antagonists, immunosuppressants, nonsteroidal antiasthmatics, anti-inflammatory agents non-steroidal, cyclooxygenase-2 inhibitors and antidiabetic agents.